1. FIELD
[0001] Provided herein are specific isoindoline compounds, pharmaceutical compositions comprising
one or more of such compounds, and methods of their use for treating, preventing,
or managing various diseases.
2. BACKGROUND
2.1 PATHOBIOLOGY OF CANCER AND OTHER DISEASES
[0002] Cancer is characterized primarily by an increase in the number of abnormal cells
derived from a given normal tissue, invasion of adjacent tissues by these abnormal
cells, or lymphatic or blood-borne spread of malignant cells to regional lymph nodes
and to distant sites (metastasis). Clinical data and molecular biologic studies indicate
that cancer is a multistep process that begins with minor preneoplastic changes, which
may under certain conditions progress to neoplasia. The neoplastic lesion may evolve
clonally and develop an increasing capacity for invasion, growth, metastasis, and
heterogeneity, especially under conditions in which the neoplastic cells escape the
host's immune surveillance.
Roitt et al., Immunology 17.1-17.12 (3rd ed., Mosby, St. Louis, Mo., 1993).
[0003] There are an enormous variety of cancers which are described in detail in the medical
literature. Examples include cancer of the lung, colon, rectum, prostate, breast,
brain, and intestine. The incidence of cancer continues to climb as the general population
ages, as new cancers develop, and as susceptible populations (
e.g., people infected with AIDS or excessively exposed to sunlight) grow. However, options
for the treatment of cancer are limited. For example, in the case of blood cancers
(
e.g., multiple myeloma), few treatment options are available, especially when conventional
chemotherapy fails and bone-marrow transplantation is not an option. A tremendous
demand therefore exists for new methods and compositions that can be used to treat
patients with cancer.
[0004] Many types of cancers are associated with new blood vessel formation, a process known
as angiogenesis. Several of the mechanisms involved in tumor-induced angiogenesis
have been elucidated. The most direct of these mechanisms is the secretion by the
tumor cells of cytokines with angiogenic properties. Examples of these cytokines include
acidic and basic fibroblastic growth factor (α,β-FGF), angiogenin, vascular endothelial
growth factor (VEGF), and TNF-α. Alternatively, tumor cells can release angiogenic
peptides through the production of proteases and the subsequent breakdown of the extracellular
matrix where some cytokines are stored (
e.g., β-FGF). Angiogenesis can also be induced indirectly through the recruitment of
inflammatory cells (particularly macrophages) and their subsequent release of angiogenic
cytokines (
e.g., TNF-α, β-FGF).
[0005] A variety of other diseases and disorders are also associated with, or characterized
by, undesired angiogenesis. For example, enhanced or unregulated angiogenesis has
been implicated in a number of diseases and medical conditions including, but not
limited to, ocular neovascular diseases, choroidal neovascular diseases, retina neovascular
diseases, rubeosis (neovascularization of the angle), viral diseases, genetic diseases,
inflammatory diseases, allergic diseases, and autoimmune diseases. Examples of such
diseases and conditions include, but are not limited to: diabetic retinopathy, retinopathy
of prematurity, corneal graft rejection, neovascular glaucoma, retrolental fibroplasia,
arthritis, and proliferative vitreoretinopathy.
[0006] Accordingly, compounds that can control angiogenesis or inhibit the production of
certain cytokines, including TNF-α, may be useful in the treatment and prevention
of various diseases and conditions.
2.2 METHODS OF TREATING CANCER
[0007] Current cancer therapy may involve surgery, chemotherapy, hormonal therapy and/or
radiation treatment to eradicate neoplastic cells in a patient (
see,
e.g., Stockdale, 1998, Medicine, vol. 3, Rubcnstcin and Federman, cds., Chapter 12, Section
IV). Recently, cancer therapy could also involve biological therapy or immunotherapy.
All of these approaches pose significant drawbacks for the patient. Surgery, for example,
may be contraindicated due to the health of a patient or may be unacceptable to the
patient. Additionally, surgery may not completely remove neoplastic tissue. Radiation
therapy is only effective when the neoplastic tissue exhibits a higher sensitivity
to radiation than normal tissue. Radiation therapy can also often elicit serious side
effects. Hormonal therapy is rarely given as a single agent. Although hormonal therapy
can be effective, it is often used to prevent or delay recurrence of cancer after
other treatments have removed the majority of cancer cells. Biological therapies and
immunotherapies are limited in number and may produce side effects such as rashes
or swellings, flu-like symptoms, including fever, chills and fatigue, digestive tract
problems, or allergic reactions.
[0009] Despite availability of a variety of chemotherapeutic agents, chemotherapy has many
drawbacks.
Stockdale, Medicine, vol. 3, Rubenstein and Federman, eds., Chapter. 12, sect. 10,
1998. Almost all chemotherapeutic agents are toxic, and chemotherapy causes significant
and often dangerous side effects including severe nausea, bone marrow depression,
and immunosuppression. Additionally, even with administration of combinations of chemotherapeutic
agents, many tumor cells are resistant or develop resistance to the chemotherapeutic
agents. In fact, those cells resistant to the particular chemotherapeutic agents used
in the treatment protocol often prove to be resistant to other drugs, even if those
agents act by different mechanism from those of the drugs used in the specific treatment.
This phenomenon is referred to as pleiotropic drug or multidrug resistance. Because
of the drug resistance, many cancers prove or become refractory to standard chemotherapeutic
treatment protocols.
[0010] Other diseases or conditions associated with, or characterized by, undesired angiogenesis
are also difficult to treat. However, some compounds such as protamine, heparin and
steroids have been proposed to be useful in the treatment of certain specific diseases.
Taylor et al., Nature 297:307 (1982);
Folkman et al., Science 221:719 (1983); and
U.S. Pat. Nos. 5,001,116 and
4,994,443.
[0011] Patent Application
WO 2008/027542 discloses 5-substituted isoindolines compounds, and pharmaceutically acceptable salts,
solvates, stereoisomers and prodrugs thereof, for use in controlling angiogenesis
or treating cancer.
[0012] Still, there is a significant need for effective methods of treating, preventing,
and managing cancer and other diseases and conditions, particularly for diseases that
are refractory to standard treatments, such as surgery, radiation therapy, chemotherapy
and hormonal therapy, while reducing or avoiding the toxicities and/or side effects
associated with the conventional therapies.
3. SUMMARY
[0013] Provided herein are specific isoindoline compounds, and pharmaceutically acceptable
salts, solvates, or stereoisomers thereof.
[0014] Specifically, in one aspect the invention provides compounds of Formula II:

or a pharmaceutically acceptable salt, solvate, or stereoisomer thereof, wherein:
X is C(=O) or CH2;
m is an integer of 0, 1, 2, or 3;
R4 is C0-4 alkyl-NR41R42; and
R41, R42 are each independently hydrogen or C1-6 alkyl.
[0015] In one embodiment, the compound of the invention, is

or a pharmaceutically acceptable salt, solvate, or stereoisomer thereof.
[0016] In another aspect, the invention provides pharmaceutical composition comprising the
compound of the invention, and one or more pharmaceutically acceptable excipients
or carriers. In one embodiment, the pharmaceutical composition of the invention further
comprises a second therapeutic agent. In another embodiment of the invention as defined
above, the composition is formulated for single dose administration. In one aspect
of any of the aforementioned embodiments, the composition is formulated as oral, parenteral,
or intravenous dosage form, optionally wherein the oral dosage form is a tablet or
capsule.
[0017] In one aspect, the compound of the invention, or the pharmaceutical composition of
the invention is for use in a method of treating, managing, or preventing a disease
or disorder, wherein the disease or disorder is cancer, a disorder associated with
angiogenesis, pain, macular degeneration or a related syndrome, a skin disease, a
pulmonary disorder, an asbestos-related disorder, a parasitic disease, an immunodeficiency
disorder, a CNS disorder, CNS injury, atherosclerosis or a related disorder, dysfunctional
sleep or a related disorder, hemoglobinopathy or a related disorder, or a TNFα related
disorder. In one embodiment, the disease is cancer. In a specific embodiment, the
cancer is hematologic or solid cancer.
[0018] In another embodiment, the compound or the pharmaceutical composition for use in
the invention further comprises the administration of one or more additional active
agents. In a specific embodiment, the use comprises the oral or parenteral administration
of the compound.
[0019] Hereinafter, reference to a/the compound or a/the pharmaceutical composition within
the context of the invention and its embodiments means the compound of Formula II
or the pharmaceutical composition comprising said compound as defined above.
[0020] Also provided herein are pharmaceutical compositions comprising a compound provided
herein,
e.g., a compound of Formula I, including a single enantiomer, a mixture of enantiomers,
or a mixture of diastereomers thereof; or a pharmaceutically acceptable salt, or solvate
thereof; in combination with one or more pharmaceutically acceptable carriers.
[0021] Further provided herein are methods of treating, preventing, or managing various
diseases in a subject, which comprise administering to the subject a therapeutically
effective amount of a compound provided herein,
e.
g., a compound of Formula I, including a single enantiomer, a mixture of enantiomers,
or a mixture of diastereomers thereof, or a pharmaceutically acceptable salt, or solvate
thereof.
4. DETAILED DESCRIPTION
4.1 DEFINITIONS
[0022] To facilitate understanding of the disclosure set forth herein, a number of terms
are defined below.
[0023] Generally, the nomenclature used herein and the laboratory procedures in organic
chemistry, medicinal chemistry, and pharmacology described herein are those well known
and commonly employed in the art. Unless defined otherwise, all technical and scientific
terms used herein generally have the same meaning as commonly understood by one of
ordinary skill in the art to which this disclosure belongs.
[0024] The term "subject" refers to an animal, including, but not limited to, a primate
(
e.g., human), cow, sheep, goat, horse, dog, cat, rabbit, rat, or mouse. The terms "subject"
and "patient" are used interchangeably herein in reference, for example, to a mammalian
subject, such as a human subject.
[0025] The terms "treat," "treating," and "treatment" refer to the eradication or amelioration
of a disease or disorder, or of one or more symptoms associated with the disease or
disorder. In general, treatment occurs after the onset of the disease or disorder.
In certain embodiments, the terms refer to minimizing the spread or worsening of the
disease or disorder resulting from the administration of one or more prophylactic
or therapeutic agents to a subject with such a disease or disorder.
[0026] The terms "prevent," "preventing," and "prevention" refer to the prevention of the
onset, recurrence or spread of a disease or disorder, or of one or more symptoms thereof.
In general, prevention occurs prior to the onset of the disease or disorder.
[0027] The terms "manage," "managing," and "management" refer to preventing or slowing the
progression, spread or worsening of a disease or disorder, or of one or more symptoms
thereof. Sometimes, the beneficial effects that a subject derives from a prophylactic
or therapeutic agent do not result in a cure of the disease or disorder.
[0028] The term "therapeutically effective amount" are meant to include the amount of a
compound that, when administered, is sufficient to prevent development of, or alleviate
to some extent, one or more of the symptoms of the disorder, disease, or condition
being treated. The term "therapeutically effective amount" also refers to the amount
of a compound that is sufficient to elicit the biological or medical response of a
cell, tissue, system, animal, or human, which is being sought by a researcher, veterinarian,
medical doctor, or clinician.
[0029] The term "IC
50" refers an amount, concentration, or dosage of a compound that is required for 50%
inhibition of a maximal response in an assay that measures such response.
[0030] The term "pharmaceutically acceptable carrier," "pharmaceutically acceptable excipient,"
"physiologically acceptable carrier," or "physiologically acceptable excipient" refers
to a pharmaceutically-acceptable material, composition, or vehicle, such as a liquid
or solid filler, diluent, excipient, solvent, or encapsulating material. In one embodiment,
each component is "pharmaceutically acceptable" in the sense of being compatible with
the other ingredients of a pharmaceutical formulation, and suitable for use in contact
with the tissue or organ of humans and animals without excessive toxicity, irritation,
allergic response, immunogenicity, or other problems or complications, commensurate
with a reasonable benefit/risk ratio.
See, Remington: The Science and Practice of Pharmacy, 21st Edition, Lippincott Williams
& Wilkins: Philadelphia, PA, 2005;
Handbook of Pharmaceutical Excipients, 5th Edition, Rowe et al., Eds., The Pharmaceutical
Press and the American Pharmaceutical Association: 2005; and
Handbook of Pharmaceutical Additives, 3rd Edition, Ash and Ash Eds., Gower Publishing
Company: 2007;
Pharmaceutical Preformulation and Formulation, Gibson Ed., CRC Press LLC: Boca Raton,
FL, 2004.
[0031] The term "about" or "approximately" means an acceptable error for a particular value
as determined by one of ordinary skill in the art, which depends in part on how the
value is measured or determined. In certain embodiments, the term "about" or "approximately"
means within 1, 2, 3, or 4 standard deviations. In certain embodiments, the term "about"
or "approximately" means within 50%, 20%, 15%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%,
1%, 0.5%, or 0.05% of a given value or range.
[0032] The terms "active ingredient" and "active substance" refer to a compound, which is
administered, alone or in combination with one or more pharmaceutically acceptable
excipients, to a subject for treating, preventing, or ameliorating one or more symptoms
of a condition, disorder, or disease. As used herein, "active ingredient" and "active
substance" may be an optically active isomer of a compound described herein.
[0033] The terms "drug," "therapeutic agent," and "chemotherapeutic agent" refer to a compound,
or a pharmaceutical composition thereof, which is administered to a subject for treating,
preventing, or ameliorating one or more symptoms of a condition, disorder, or disease.
[0034] The term "alkyl" refers to a linear or branched saturated monovalent hydrocarbon
radical, wherein the alkyl may optionally be substituted with one or more substituents.
The term "alkyl" also encompasses both linear and branched alkyl, unless otherwise
specified. In certain embodiments, the alkyl is a linear saturated monovalent hydrocarbon
radical that has 1 to 20 (C
1-20), 1 to 15 (C
1-15), 1 to 12 (C
1-12), 1 to 10 (C
1-10), or 1 to 6 (C
1-6) carbon atoms, or branched saturated monovalent hydrocarbon radical of 3 to 20 (C
3-20), 3 to 15 (C
3-15), 3 to 12 (C
3-12), 3 to 10 (C
3-10), or 3 to 6 (C
3-6) carbon atoms. As used herein, linear C
1-6 and branched C
3-6 alkyl groups are also referred as "lower alkyl." Examples of alkyl groups include,
but are not limited to, methyl, ethyl, propyl (including all isomeric forms), n-propyl,
isopropyl, butyl (including all isomeric forms), n-butyl, isobutyl, t-butyl, pentyl
(including all isomeric forms), and hexyl (including all isomeric forms). For example,
C
1-6 alkyl refers to a linear saturated monovalent hydrocarbon radical of 1 to 6 carbon
atoms or a branched saturated monovalent hydrocarbon radical of 3 to 6 carbon atoms.
[0035] The term "alkenyl" refers to a linear or branched monovalent hydrocarbon radical,
which contains one or more, in one embodiment, one to five, carbon-carbon double bonds.
The alkenyl may be optionally substituted one or more substituents. The term "alkenyl"
also embraces radicals having "
cis" and "
trans" configurations, or alternatively, "E" and "Z" configurations, as appreciated by
those of ordinary skill in the art. As used herein, the term "alkenyl" encompasses
both linear and branched alkenyl, unless otherwise specified. For example, C
2-6 alkenyl refers to a linear unsaturated monovalent hydrocarbon radical of 2 to 6 carbon
atoms or a branched unsaturated monovalent hydrocarbon radical of 3 to 6 carbon atoms.
In certain embodiments, the alkenyl is a linear monovalent hydrocarbon radical of
2 to 20 (C
2-20), 2 to 15 (C
2-15), 2 to 12 (C
2-12), 2 to 10 (C
2-10), or 2 to 6 (C
2-6) carbon atoms, or a branched monovalent hydrocarbon radical of 3 to 20 (C
3-20), 3 to 15 (C
3-15), 3 to 12 (C
3-12), 3 to 10 (C
3-10), or 3 to 6 (C
3-6) carbon atoms. Examples of alkenyl groups include, but are not limited to, ethenyl,
propen-1-yl, propen-2-yl, allyl, butenyl, and 4-methylbutenyl.
[0036] The term "alkynyl" refers to a linear or branched monovalent hydrocarbon radical,
which contains one or more, in one embodiment, one to five, carbon-carbon triple bonds.
The alkynyl may be optionally substituted one or more substituents. The term "alkynyl"
also encompasses both linear and branched alkynyl, unless otherwise specified. In
certain embodiments, the alkynyl is a linear monovalent hydrocarbon radical of 2 to
20 (C
2-20), 2 to 15 (C
2-15), 2 to 12 (C
2-12), 2 to 10 (C
2-10), or 2 to 6 (C
2-6) carbon atoms, or a branched monovalent hydrocarbon radical of 3 to 20 (C
3-20), 3 to 15 (C
3-15), 3 to 12 (C
3-12), 3 to 10 (C
3-10), or 3 to 6 (C
3-6) carbon atoms. Examples of alkynyl groups include, but are not limited to, ethynyl
(-C≡CH) and propargyl (-CH
2C≡CH). For example, C
2-6 alkynyl refers to a linear unsaturated monovalent hydrocarbon radical of 2 to 6 carbon
atoms or a branched unsaturated monovalent hydrocarbon radical of 3 to 6 carbon atoms.
[0037] The term "cycloalkyl" refers to a cyclic saturated bridged and/or non-bridged monovalent
hydrocarbon radical, which may be optionally substituted one or more substituents.
In certain embodiments, the cycloalkyl has from 3 to 20 (C
3-20), from 3 to 15 (C
3-15), from 3 to 12 (C
3-12), from 3 to 10 (C
3-10), or from 3 to 7 (C
3-7) carbon atoms. Examples of cycloalkyl groups include, but are not limited to, cyclopropyl,
cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, decalinyl, and adamantyl.
[0038] The term "aryl" refers to a monocyclic aromatic group and/or multicyclic monovalent
aromatic group that contain at least one aromatic hydrocarbon ring. In certain embodiments,
the aryl has from 6 to 20 (C
6-20), from 6 to 15 (C
6-15), or from 6 to 10 (C
6-10) ring atoms. Examples of aryl groups include, but are not limited to, phenyl, naphthyl,
fluorenyl, azulenyl, anthryl, phenanthryl, pyrenyl, biphenyl, and terphenyl. Aryl
also refers to bicyclic or tricyclic carbon rings, where one of the rings is aromatic
and the others of which may be saturated, partially unsaturated, or aromatic, for
example, dihydronaphthyl, indenyl, indanyl, or tetrahydronaphthyl (tetralinyl). In
certain embodiments, aryl may also be optionally substituted with one or more substituents.
[0039] The term "aralkyl" or "aryl-alkyl" refers to a monovalent alkyl group substituted
with aryl. In certain embodiments, both alkyl and aryl may be optionally substituted
with one or more substituents.
[0040] The term "heteroaryl" refers to a monocyclic aromatic group and/or multicyclic aromatic
group that contain at least one aromatic ring, wherein at least one aromatic ring
contains one or more heteroatoms independently selected from O, S, and N. Each ring
of a heteroaryl group can contain one or two O atoms, one or two S atoms, and/or one
to four N atoms, provided that the total number of heteroatoms in each ring is four
or less and each ring contains at least one carbon atom. The heteroaryl may be attached
to the main structure at any heteroatom or carbon atom which results in the creation
of a stable compound. In certain embodiments, the heteroaryl has from 5 to 20, from
5 to 15, or from 5 to 10 ring atoms. Examples of monocyclic heteroaryl groups include,
but are not limited to, pyrrolyl, pyrazolyl, pyrazolinyl, imidazolyl, oxazolyl, isoxazolyl,
thiazolyl, thiadiazolyl, isothiazolyl, furanyl, thienyl, oxadiazolyl, pyridyl, pyrazinyl,
pyrimidinyl, pyridazinyl, and triazinyl. Examples of bicyclic heteroaryl groups include,
but are not limited to, indolyl, benzothiazolyl, benzoxazolyl, benzothienyl, quinolinyl,
tetrahydroisoquinolinyl, isoquinolinyl, benzimidazolyl, benzopyranyl, indolizinyl,
benzofuranyl, isobenzofuranyl, chromonyl, coumarinyl, cinnolinyl, quinoxalinyl, indazolyl,
purinyl, pyrrolopyridinyl, furopyridinyl, thienopyridinyl, dihydroisoindolyl, and
tetrahydroquinolinyl. Examples of tricyclic heteroaryl groups include, but are not
limited to, carbazolyl, benzindolyl, phenanthrollinyl, acridinyl, phenanthridinyl,
and xanthenyl. In certain embodiments, heteroaryl may also be optionally substituted
with one or more substituents.
[0041] The term "heterocyclyl" or "heterocyclic" refers to a monocyclic non-aromatic ring
system and/or multicyclic ring system that contains at least one non-aromatic ring,
wherein one or more of the non-aromatic ring atoms are heteroatoms independently selected
from O, S, or N; and the remaining ring atoms are carbon atoms. In certain embodiments,
the heterocyclyl or heterocyclic group has from 3 to 20, from 3 to 15, from 3 to 10,
from 3 to 8, from 4 to 7, or from 5 to 6 ring atoms. In certain embodiments, the heterocyclyl
is a monocyclic, bicyclic, tricyclic, or tetracyclic ring system, which may includes
a fused or bridged ring system, and in which the nitrogen or sulfur atoms may be optionally
oxidized, the nitrogen atoms may be optionally quaternized, and some rings may be
partially or fully saturated, or aromatic. The heterocyclyl may be attached to the
main structure at any heteroatom or carbon atom which results in the creation of a
stable compound. Examples of such heterocyclic radicals include, but are not limited
to, acridinyl, azepinyl, benzimidazolyl, benzindolyl, benzoisoxazolyl, benzisoxazinyl,
benzodioxanyl, benzodioxolyl, benzofuranonyl, benzofuranyl, benzonaphthofuranyl, benzopyranonyl,
benzopyranyl, benzotetrahydrofuranyl, benzotetrahydrothienyl, benzothiadiazolyl, benzothiazolyl,
benzothiophenyl, benzotriazolyl, benzothiopyranyl, benzoxazinyl, benzoxazolyl, benzothiazolyl,
β-carbolinyl, carbazolyl, chromanyl, chromonyl, cinnolinyl, coumarinyl, decahydroisoquinolinyl,
dibenzofuranyl, dihydrobenzisothiazinyl, dihydrobenzisoxazinyl, dihydrofuryl, dihydropyranyl,
dioxolanyl, dihydropyrazinyl, dihydropyridinyl, dihydropyrazolyl, dihydropyrimidinyl,
dihydropyrrolyl, dioxolanyl, 1,4-dithianyl, furanonyl, furanyl, imidazolidinyl, imidazolinyl,
imidazolyl, imidazopyridinyl, imidazothiazolyl, indazolyl, indolinyl, indolizinyl,
indolyl, isobenzotetrahydrofuranyl, isobenzotetrahydrothienyl, isobenzothienyl, isochromanyl,
isocoumarinyl, isoindolinyl, isoindolyl, isoquinolinyl, isothiazolidinyl, isothiazolyl,
isoxazolidinyl, isoxazolyl, morpholinyl, naphthyridinyl, octahydroindolyl, octahydroisoindolyl,
oxadiazolyl, oxazolidinonyl, oxazolidinyl, oxazolopyridinyl, oxazolyl, oxiranyl, perimidinyl,
phenanthridinyl, phenathrolinyl, phenarsazinyl, phenazinyl, phenothiazinyl, phenoxazinyl,
phthalazinyl, piperazinyl, piperidinyl, 4-piperidonyl, pteridinyl, purinyl, pyrazinyl,
pyrazolidinyl, pyrazolyl, pyridazinyl, pyridinyl, pyridopyridinyl, pyrimidinyl, pyrrolidinyl,
pyrrolinyl, pyrrolyl, quinazolinyl, quinolinyl, quinoxalinyl, quinuclidinyl, tetrahydrofuryl,
tetrahydrofuranyl, tetrahydroisoquinolinyl, tetrahydropyranyl, tetrahydrothienyl,
tetrazolyl, thiadiazolopyrimidinyl, thiadiazolyl, thiamorpholinyl, thiazolidinyl,
thiazolyl, thienyl, triazinyl, triazolyl, and 1,3,5-trithianyl. In certain embodiments,
heterocyclic may also be optionally substituted with one or more substituents.
[0042] The term "halogen", "halide" or "halo" refers to fluorine, chlorine, bromine, and/or
iodine.
[0043] The terms "optically active" and "enantiomerically active" refer to a collection
of molecules, which has an enantiomeric excess of no less than about 50%, no less
than about 70%, no less than about 80%, no less than about 90%, no less than about
91%, no less than about 92%, no less than about 93%, no less than about 94%, no less
than about 95%, no less than about 96%, no less than about 97%, no less than about
98%, no less than about 99%, no less than about 99.5%, or no less than about 99.8%.
[0044] In describing an optically active compound, the prefixes R and S arc used to denote
the absolute configuration of the molecule about its chiral center(s). The (+) and
(-) are used to denote the optical rotation of the compound, that is, the direction
in which a plane of polarized light is rotated by the optically active compound. The
(-) prefix indicates that the compound is levorotatory, that is, the compound rotates
the plane of polarized light to the left or counterclockwise. The (+) prefix indicates
that the compound is dextrorotatory, that is, the compound rotates the plane of polarized
light to the right or clockwise. However, the sign of optical rotation, (+) and (-),
is not related to the absolute configuration of the molecule, R and S.
[0045] The term "solvate" refers to a compound provided herein or a salt thereof, which
further includes a stoichiometric or non-stoichiometric amount of solvent bound by
non-covalent intermolecular forces. Where the solvent is water, the solvate is a hydrate.
4.2 COMPOUNDS
[0046] Provided herein are specific isoindoline compounds, pharmaceutical compositions comprising
one or more of such compounds, and methods of their use for treating, preventing,
or managing various diseases.
[0047] Specifically, in one aspect the invention provides compounds of Formula II:

or a pharmaceutically acceptable salt, solvate, or stereoisomer thereof, wherein:
X is C(=O) or CH2;
m is an integer of 0, 1, 2, or 3;
R4 is C0-4 alkyl-NR41R42; and
R41, R42 are each independently hydrogen or C1-6 alkyl.
[0048] In one embodiment, the compound of the invention, is

or a pharmaceutically acceptable salt, solvate, or stereoisomer thereof.
[0049] In another aspect, the invention provides pharmaceutical composition comprising the
compound of the invention, and one or more pharmaceutically acceptable excipients
or carriers. In one embodiment, the pharmaceutical composition of the invention further
comprises a second therapeutic agent. In another embodiment of the invention as defined
above, the composition is formulated for single dose administration. In one aspect
of any of the aforementioned embodiments, the composition is formulated as oral, parenteral,
or intravenous dosage form, optionally wherein the oral dosage form is a tablet or
capsule.
[0050] In one aspect, the compound of the invention, or the pharmaceutical composition of
the invention is for use in a method of treating, managing, or preventing a disease
or disorder, wherein the disease or disorder is cancer, a disorder associated with
angiogenesis, pain, macular degeneration or a related syndrome, a skin disease, a
pulmonary disorder, an asbestos-related disorder, a parasitic disease, an immunodeficiency
disorder, a CNS disorder, CNS injury, atherosclerosis or a related disorder, dysfunctional
sleep or a related disorder, hemoglobinopathy or a related disorder, or a TNFα related
disorder. In one embodiment, the disease is cancer. In a specific embodiment, the
cancer is hematologic or solid cancer.
[0051] In another embodiment, the compound or the pharmaceutical composition for use in
the invention further comprises the administration of one or more additional active
agents. In a specific embodiment, the use comprises the oral or parenteral administration
of the compound.
[0052] For reference only, disclosed herein is a compound of Formula I:

or a pharmaceutically acceptable salt, solvate, prodrug, or stereoisomer thereof,
wherein:
X is C(=O) or CH2;
Y is O, cyanamido (N-≡N), or amido (NH);
m is an integer of 0, 1, 2, or 3;
R1 is hydrogen or C1-6 alkyl;
R2 is hydrogen, -NO2, C1-10 alkyl, C0-6 alkyl-(5 to 10 membered heteroaryl), C0-6 alkyl-(5 to 6 membered heterocyclyl), C0-6 alkyl-OH, C0-4 alkyl-NH2, -NHCO-C1-6 alkyl, -OR21, or-(CH2-Z)0-2-(5 to 10 membered heteroaryl), where each heteroaryl and heterocyclyl is optionally
substituted with one or more C1-6 alkyl;
R3 is hydrogen, halogen, -NO2, C0-6 alkyl-(5 to 10 membered heteroaryl), C0-6 alkyl-(5 to 6 membered heterocyclyl), C0-6 alkyl-OH, C0-4 alkyl-NH2, -NHCO-C1-6 alkyl, -OR21, or -(CH2-Z)0-2-(5 to 10 membered heteroaryl), where each heteroaryl and heterocyclyl is optionally
substituted with one or more C1-6 alkyl;
R21 is C6-10 aryl, 5 to 10 membered heteroaryl, 5 to 6 membered heterocyclyl, or - CO(CH2)0-2R22, wherein the aryl, heteroaryl, and heterocyclyl are each optionally substituted with
one or more C1-6 alkyl;
R22 is -NH2 or 5 to 6 membered heterocyclyl; and
Z is CH2, NH, or O;
with the proviso that when R1 is hydrogen, then R2 is not hydrogen or C1-10 alkyl;
with the proviso that when Y is O, then R3 is not halogen; and
with the proviso that when Y is O and R3 is halogen, then R2 is C0-6 alkyl-(5-6 membered heterocyclyl).
[0053] In certain examples, X is CH
2. In certain examples, X is C(=O).
[0054] In certain examples, Y is O. In certain examples, Y is cyanamido. In certain examples,
Y is amido.
[0055] In certain examples, Z is CH
2. In certain examples, Z is NH. In certain examples, Z is O.
[0056] In certain examples, m is 0. In certain examples, m is 1. In certain examples, m
is 2. In certain examples, m is 3.
[0057] In certain examples, R
1 is hydrogen. In certain examples, R
1 is C
1-6 alkyl, optionally substituted with one, two, or three substituents Q as described
herein. In certain examples, R is methyl.
[0058] In certain examples, R
2 is hydrogen. In certain examples, R
2 is halogen. In certain examples, R
2 is nitro. In certain examples, R
2 is C
1-10 alkyl. In certain examples, R
2 is C
0-6 alkyl-(5 to 10 membered heteroaryl), where the heteroaryl is optionally substituted
with one or more C
1-6 alkyl. In certain examples, R
2 is C
0-6 alkyl-(5 to 6 membered heterocyclyl), where the heterocyclyl is optionally substituted
with one or more C
1-6 alkyl. In certain examples, R
2 is C
0-6 alkyl-OH. In certain examples, R
2 is C
0-4 alkyl-NH
2. In certain examples, R
2 is -NHCO-C
1-6 alkyl. In certain examples, R
2 is -OR
21, wherein R
21 is as described herein. In certain examples, R
2 is or -(CH
2-Y)
0-2-(5 to 10 membered heteroaryl), where the heteroaryl is optionally substituted with
one or more C
1-6 alkyl. In certain examples, R
2 is hydrogen, amino, acetamido, hydroxy, nitro, aminomethyl, hydroxymethyl, 2-methyl-1
H-imidazol-1-yl, 3-methyl-1,2,4-oxadiazol-5-yl, 4-methylpiperazin-1-yl)methyl, 2-methyl-2
H-pyrazol-3-yl, 1-methyl-1
H-pyrazol-3-yl, 2-methylthiazol-4-yl, 4-methyl-4
H-1,2,4-triazol-3-yl, morpholinomethyl, (pyridin-4-yl)methyl, (pyridin-4-yloxy)methyl,
pheoxy, pyridin-2-yloxy, piperidin-4-yloxy, 2-aminoacetoxy, or 2-piperazin-1-ylacetoxy.
[0059] In certain examples, R
3 is hydrogen. In certain examples, R
3 is nitro. In certain examples, R
3 is C
0-6 alkyl-(5 to 10 membered heteroaryl), where the heteroaryl is optionally substituted
with one or more C
1-6 alkyl. In certain examples, R
3 is C
0-6 alkyl-(5 to 6 membered heterocyclyl), where the heterocyclyl is optionally substituted
with one or more C
1-6 alkyl. In certain examples, R
3 is C
0-6 alkyl-OH. In certain examples, R
3 is C
0-4 alkyl-NH
2- In certain examples, is -NHCO-C
1-6 alkyl. In certain examples, R
3 is -OR
21, wherein R
21 is as described herein. In certain examples, R
3 is or -(CH
2-Y)
0-2-(5 to 10 membered heteroaryl), where the heteroaryl is optionally substituted with
one or more C
1-6 alkyl. In certain examples, R
3 is hydrogen, amino, acetamido, hydroxy, nitro, methyl, aminomethyl, hydroxymethyl,
2-methyl-1
H-imidazol-1-yl, 3-methyl-1,2,4-oxadiazol-5-yl, 4-methylpiperazin-1-yl)methyl, 2-methyl-2
H-pyrazol-3-yl, 1-methyl-1
H-pyrazol-3-yl, 2-methylthiazol-4-yl, 4-methyl-4
H-1,2,4-triazol-3-yl, morpholinomethyl, (pyridin-4-yl)methyl, (pyridin-4-yloxy)methyl,
pheoxy, pyridin-2-yloxy, piperidin-4-yloxy, 2-aminoacetoxy, or 2-piperazin-1-ylacetoxy.
[0061] In one aspect of the invention, provided herein is a compound of Formula II:

or a pharmaceutically acceptable salt, solvate, or stereoisomer thereof, wherein:
X is C(=O) or CH2;
m is an integer of 0, 1, 2, or 3;
R4 is C0-4 alkyl-NR41R42; and
R41, R42 are each independently hydrogen or C1-6 alkyl.
[0062] In certain embodiments, X is CH
2. In certain embodiments, X is C(=O).
[0063] In certain embodiments, m is 0. In certain embodiments, m is 1. In certain embodiments,
m is 2. In certain embodiments, m is 3.
[0064] In some cases, R
4 is C
3-10 cycloalkyl, optionally substituted with one or more (C
1-6) alkyl or C
0-4 alkyl-C
6-10 aryl. In other cases, R
4 is 5 to 6 membered heterocyclyl, optionally substituted with one or more (C
1-6) alkyl or C
0-4 alkyl-C
6-10 aryl. According to the invention, R
4 is C
0-4 alkyl-NR
41 R
42, wherein R
41 and R
42 are each described herein.
[0065] In certain embodiments, R
4 is 3-(
N,
N-diethylamino)propyl. In other examples disclosed herein, R4 is 4-acetamidophenyl,
3-(2-aminoacctoxy)-4-mcthylphcnyl, 3-aminomethyl-4-methylphenyl, 2-aminomethyl-5-methylphenyl,
3-aminophenyl, 3-amino-4-methylphenyl, 3-chloro-4-methylphenyl, 4-hydroxymethylphenyl,
3-hydroxy-4-methylphenyl, 3-(2-methyl-1
H-imidazol-1-yl)phenyl, 4-methyl-3-nitrophenyl, 3-(3-methyl-1,2,4-oxadiazol-5-yl)phenyl,
4-methyl-3-(2-piperazin-1-ylacetoxy)-phenyl, 3-((4-methylpiperazin-1-yl)methyl)phenyl,
3-(1-methyl-1
H-pyrazol-3-yl)phenyl, 3-(2-methyl-2
H-pyrazol-3-yl)phenyl, 3-(2-methylthiazol-4-yl)phenyl, 4-(4-methyl-4
H-1,2,4-triazol-3-yl)phenyl, 3-(morpholinomethyl)phenyl, 4-(morpholinomethyl)phenyl,
4-nitrophenyl, phenyl, 3-(piperidin-4-yloxy)phenyl, 4-(pyridin-4-yl)methylphenyl,
4-((pyridin-4-yloxy)methyl)phenyl, 3-(pyridin-2-yloxy)phenyl, 3-phenoxyphenyl, 4-
tert-butylcyclohexyl,
cis-4-
tert-butylcyclohexyl,
trans-4-
tert-butylcyclohexyl, 4-methylcyclohexyl,
cis-4-methylcyclohexyl,
trans-4-methylcyclohexyl, 1-benzylpiperidin-4-yl, 4-methyltetrahydro-2
H-pyran-4-yl, piperidin-4-yl, 4-phenylcyclohexyl,
cis-4-phenylcyclohexyl, or
trans-4-phenylcyclohexyl.
[0066] In one embodiment, the compound is:

or a pharmaceutically acceptable salt, solvate, or stereoisomer thereof.
[0067] In other examples disclosed herein is a compound of Formula III:

or a pharmaceutically acceptable salt, solvate, prodrug, or stereoisomer thereof,
wherein:
X is C(=O) or CH2;
m is an integer of 0, 1, 2, or 3;
R5 and R6 are each independently: hydrogen, halo, C1-6 alkyl, oxo, -NO2, C1-6 alkoxy, -Z-C1-6 alkyl, , C0-6 alkyl-(5 to 10 membered heteroaryl), C0-6 alkyl-(5 to 6 membered heterocyclyl), C0-6 alkyl-OH, C0-4 alkyl-NH2, -NHCO-C1-6 alkyl, -OR21, or -(CH2-Y)0-2-(5 to 10 membered heteroaryl),
wherein Z is S or SO2;
wherein R21 is as defined above;
wherein each heteroaryl and heterocyclyl above is optionally substituted with one
or more C1-6 alkyl; and
wherein the alkyl or alkoxy above may be optionally substituted with one or more:
halogen; cyano; nitro; amino; C1-6 alkylidenedioxy; C1-6 alkoxy, itself optionally substituted with one or more halogens; or C1-6 alkylthio, itself optionally substituted with one or more halogens;
R7 is -COR71 or -PO(OR72)(OR73);
R71 is C1-10 alkyl, C6-10 aryl, or 5 to 6 membered heterocyclyl; wherein the alkyl, aryl, heterocyclyl may
be optionally substituted with one or more amino, C1-6 alkylamino, di(C1-6 alkyl)amino, or -COOR74; and
R72, R73, and R74 are ach independently hydrogen or C1-10 alkyl.
[0068] In certain examples, X is CH
2. In certain examples, X is C(=O).
[0069] In certain examples, m is 0. In certain examples, m is 1. In certain examples, m
is 2. In certain examples, m is 3.
[0070] In certain examples, R
5 is hydrogen. In certain examples, R
5 is halo. In certain examples, R
5 is fluoro or chloro.
[0071] In certain examples, R
6 is hydrogen. In certain examples, R
6 is halo. In certain examples, R
6 is fluoro or chloro.
[0072] In certain examples, R
7 is -COR
41, wherein R
41 is as described herein. In certain examples, R
7 is -PO(OR
42))(OR
43), wherein R and R are each as described herein.
[0074] In yet another example, disclosed herein is a compound of Formula IV:

or a pharmaceutically acceptable salt, solvate, prodrug, or stereoisomer thereof,
wherein:
X is C(=O) or CH2;
n is an integer of 0 or 1;
R8 is hydrogen or halo; and
R9 is hydrogen, amino, or 5 to 10 membered heteroaryl or heterocyclyl;
with the proviso that when m is 0, R9 is not hydrogen.
[0075] In certain examples, X is CH
2. In certain examples, X is C(=O).
[0076] In certain examples, n is 0. In certain examples, n is 1.
[0077] In certain examples, R
8 is hydrogen. In certain examples, R
8 is halo. In certain examples, R
8 is fluoro or chloro.
[0078] In certain examples, R
9 is hydrogen. In certain examples, R
9 is amino. In certain examples, R
9 is 5 to 10 membered heteroaryl. In certain examples, R
9 is 5 to 10 membered heterocyclyl.
[0079] For example, the compound is:

or a pharmaceutically acceptable salt, solvate, prodrug, or stereoisomer thereof.
[0080] In yet another examples, disclosed herein is a compound of Formula V:

or a pharmaceutically acceptable salt, solvate, prodrug, or stereoisomer thereof,
wherein:
X is C(=O) or CH2;
m is an integer of 0, 1, 2, or 3;
R10 and R11 are each independently hydrogen, halo, C1-6 alkyl, or C6-10 aryloxy, wherein the alkyl and aryl are each optionally substituted with one or more
halo.
[0081] In certain examples, X is CH
2. In certain examples, X is C(=O).
[0082] In certain examples, m is 0. In certain examples, m is 1. In certain examples, m
is 2. In certain examples, m is 3.
[0083] In certain examples, R
10 is hydrogen. In certain examples, R is halo. In certain examples, R
10 is fluoro or chloro. In certain examples, R
10 is C
1-6 alkyl, optionally substituted with one or more halo. In certain examples, R
10 is C
6-10 aryloxy, optionally substituted with one or more halo.
[0084] In certain examples, R
11 is hydrogen. In certain examples, R
11 is halo. In certain examples, R
11 is fluoro or chloro. In certain examples, R
11 is C
1-6 alkyl, optionally substituted with one or more halo. In certain examples, R
11 is C
6-10 aryloxy, optionally substituted with one or more halo.
[0086] In yet another example, disclosed herein is a compound of Formula VI:

or a pharmaceutically acceptable salt, solvate, prodrug, or stereoisomer thereof,
wherein:
X is CH2 or C=O
m and n are each independently 0 or 1;
p is 0, 1, 2, or 3;
R81 is 5 to 6 membered heterocyclyl, optionally substituted with C1-6 alkyl; and
R82 is hydrogen or halogen.
[0087] In one example, X is CH
2- In another example, X is C=O.
[0088] In one example, m is 0. In another example, m is 1. In another example, n is 0. In
another example, n is 1.
[0089] In one example, p is 0. In another example, p is 1. In another example, p is 2. In
another example, p is 3
.
[0090] In example, R
81 is 5 membered heterocycle. In another example, the 5 membered heterocycle is substituted
with C
1-6 alkyl. In another example, R
81 is 6 membered heterocycle. In another example, the 6 membered heterocycle is substituted
with C
1-6 alkyl.
[0091] In one example, R
82 is hydrogen. In another example, R
82 is halogen.
[0093] In yet another example, disclosed herein is a compound of the following formula:

or a pharmaceutically acceptable salt, solvate, prodrug, or stereoisomer thereof.
[0094] The compounds provided herein are intended to encompass all possible stereoisomers,
unless a particular stereochemistry is specified. Where the compound provided herein
contains an alkenyl or alkenylene group, the compound may exist as one or mixture
of geometric
cis/
trans (or Z/E) isomers. Where structural isomers are interconvertible
via a low energy barrier, the compound may exist as a single tautomer or a mixture of
tautomers. This can take the form of proton tautomerism in the compound that contains,
for example, an imino, keto, or oxime group; or so-called valence tautomerism in the
compound that contain an aromatic moiety. It follows that a single compound may exhibit
more than one type of isomerism.
[0095] The compounds provided herein may be enantiomerically pure, such as a single enantiomer
or a single diastereomer, or be stereoisomeric mixtures, such as a mixture of enantiomers,
a racemic mixture, or a diastereomeric mixture. As such, one of skill in the art will
recognize that administration of a compound in its (
R) form is equivalent, for compounds that undergo epimerization
in vivo, to administration of the compound in its (
S) form. Conventional techniques for the preparation/isolation of individual enantiomers
include synthesis from a suitable optically pure precursor, asymmetric synthesis from
achiral starting materials, or resolution of an enantiomeric mixture, for example,
chiral chromatography, recrystallization, resolution, diastereomeric salt formation,
or derivatization into diastereomeric adducts followed by separation.
[0096] When the compound provided herein contains an acidic or basic moiety, it may also
be provided as a pharmaceutically acceptable salt (
See,
Berge et al., J. Pharm. Sci. 1977, 66, 1-19; and "
Handbook of Pharmaceutical Salts, Properties, and Use," Stahl and Wermuth, Ed.; Wiley-VCH
and VHCA, Zurich, 2002).
[0097] Suitable acids for use in the preparation of pharmaceutically acceptable salts include,
but are not limited to, acetic acid, 2,2-dichloroacetic acid, acylated amino acids,
adipic acid, alginic acid, ascorbic acid, L-aspartic acid, benzenesulfonic acid, benzoic
acid, 4-acetamidobenzoic acid, boric acid, (+)-camphoric acid, camphorsulfonic acid,
(+)-(1
S)-camphor-10-sulfonic acid, capric acid, caproic acid, caprylic acid, cinnamic acid,
citric acid, cyclamic acid, cyclohexanesulfamic acid, dodecylsulfuric acid, ethane-1,2-disulfonic
acid, ethanesulfonic acid, 2-hydroxy-ethanesulfonic acid, formic acid, fumaric acid,
galactaric acid, gentisic acid, glucoheptonic acid, D-gluconic acid, D-glucuronic
acid, L-glutamic acid, α-oxoglutaric acid, glycolic acid, hippuric acid, hydrobromic
acid, hydrochloric acid, hydroiodic acid, (+)-L-lactic acid, (±)-DL-lactic acid, lactobionic
acid, lauric acid, maleic acid, (-)-L-malic acid, malonic acid, (±)-DL-mandelic acid,
methanesulfonic acid, naphthalene-2-sulfonic acid, naphthalene-1,5-disulfonic acid,
1-hydroxy-2-naphthoic acid, nicotinic acid, nitric acid, oleic acid, orotic acid,
oxalic acid, palmitic acid, pamoic acid, perchloric acid, phosphoric acid, L-pyroglutamic
acid, saccharic acid, salicylic acid, 4-amino-salicylic acid, sebacic acid, stearic
acid, succinic acid, sulfuric acid, tannic acid, (+)-L-tartaric acid, thiocyanic acid,
p-toluenesulfonic acid, undecylenic acid, and valeric acid.
[0098] Suitable bases for use in the preparation of pharmaceutically acceptable salts, including,
but not limited to, inorganic bases, such as magnesium hydroxide, calcium hydroxide,
potassium hydroxide, zinc hydroxide, or sodium hydroxide; and organic bases, such
as primary, secondary, tertiary, and quaternary, aliphatic and aromatic amines, including
L-arginine, benethamine, benzathine, choline, deanol, diethanolamine, diethylamine,
dimethylamine, dipropylamine, diisopropylamine, 2-(diethylamino)-ethanol, ethanolamine,
ethylamine, ethylenediamine, isopropylamine, N-methyl-glucamine, hydrabamine, 1H-imidazole,
L-lysine, morpholine, 4-(2-hydroxyethyl)-morpholine, methylamine, piperidine, piperazine,
propylamine, pyrrolidine, 1-(2-hydroxyethyl)-pyrrolidine, pyridine, quinuclidine,
quinoline, isoquinoline, secondary amines, triethanolamine, trimethylamine, triethylamine,
N-methyl-D-glucamine, 2-amino-2-(hydroxymethyl)-1,3-propanediol, and tromethamine.
[0099] The compound disclosed herein may also be provided as a prodrug, which is a functional
derivative of the compound and is readily convertible into the parent compound in
vivo. Prodrugs are often useful because, in some situations, they may be easier to
administer than the parent compound. They may, for instance, be bioavailable by oral
administration whereas the parent compound is not. The prodrug may also have enhanced
solubility in pharmaceutical compositions over the parent compound. A prodrug may
be converted into the parent drug by various mechanisms, including enzymatic processes
and metabolic hydrolysis. See
Harper, Progress in Drug Research 1962, 4, 221-294;
Morozowich et al. in "Design of Biopharmaceutical Properties through Prodrugs and
Analogs," Roche Ed., APHA Acad. Pharm. Sci. 1977; "
Bioreversible Carriers in Drug in Drug Design, Theory and Application," Roche Ed.,
APHA Acad. Pharm. Sci. 1987; "
Design of Prodrugs," Bundgaard, Elsevier, 1985;
Wang et al., Curr. Pharm. Design 1999, 5, 265-287;
Pauletti et al., Adv. Drug. Delivery Rev. 1997, 27, 235-256;
Mizen et al., Pharm. Biotech. 1998, 11, 345-365;
Gaignault et al., Pract. Med. Chem. 1996, 671-696; Asgharnejad in "
Transport Processes in Pharmaceutical Systems," Amidon et al., Ed., Marcell Dekker,
185-218, 2000;
Balant et al., Eur. J. Drug Metab. Pharmacokinet. 1990, 15, 143-53;
Balimane and Sinko, Adv. Drug Delivery Rev. 1999, 39, 183-209;
Browne, Clin. Neuropharmacol. 1997, 20, 1-12;
Bundgaard, Arch. Pharm. Chem. 1979, 86, 1-39;
Bundgaard, Controlled Drug Delivery 1987, 17, 179-96;
Bundgaard, Adv. Drug Delivery Rev. 1992, 8, 1-38;
Fleisher et al., Adv. Drug Delivery Rev. 1996, 19, 115-130;
Fleisher et al., Methods Enzymol. 1985, 112, 360-381;
Farquhar et al., J. Pharm. Sci. 1983, 72, 324-325;
Freeman et al., J. Chem. Soc., Chem. Commun. 1991, 875-877;
Friis and Bundgaard, Eur. J. Pharm. Sci. 1996, 4, 49-59;
Gangwar et al., Des. Biopharm. Prop. Prodrugs Analogs, 1977, 409-421;
Nathwani and Wood, Drugs 1993, 45, 866-94;
Sinhababu and Thakker, Adv. Drug Delivery Rev. 1996, 19, 241-273;
Stella et al., Drugs 1985, 29, 455-73;
Tan et al., Adv. Drug Delivery Rev. 1999, 39, 117-151;
Taylor, Adv. Drug Delivery Rev. 1996, 19, 131-148;
Valentino and Borchardt, Drug Discovery Today 1997, 2, 148-155;
Wiebe and Knaus, Adv. Drug Delivery Rev. 1999, 39, 63-80; and
Waller et al., Br. J. Clin. Pharmac. 1989, 28, 497-507.
4.3 COMPOUNDS AND PHARMACEUTICAL COMPOSITIONS FOR USE IN METHODS OF TREATMENT, PREVENTION AND MANAGEMENT
[0100] In one embodiment, provided herein are compounds and pharmaceutical compositions
of the invention for use in methods of treating, preventing, and/or managing various
diseases or disorders wherein said compounds and pharmaceutical composition include
a single enantiomer, a mixture of enantiomers, or a mixture of diastereomers thereof,
or a pharmaceutically acceptable salt, or solvate, thereof. Without being limited
by a particular theory, compounds provided herein can control angiogenesis or inhibit
the production of certain cytokines including, but not limited to, TNF-α, IL-1β, IL-12,
IL-18, GM-CSF, and/or IL-6. Without being limited by a particular theory, compounds
provided herein can stimulate the production of certain other cytokines including
IL-10, and also act as a costimulatory signal for T cell activation, resulting in
increased production of cytokines such as, but not limited to, IL-12 and/or IFN-γ.
In addition, compounds provided herein can enhance the effects of NK cells and antibody-mediated
cellular cytotoxicity (ADCC). Further, compounds provided herein may be immunomodulatory
and/or cytotoxic, and thus, may be useful as chemotherapeutic agents. Consequently,
without being limited by a particular theory, some or all of such characteristics
possessed by the compounds provided herein may render them useful in treating, managing,
and/or preventing various diseases or disorders. In one embodiment, the compounds
provided herein are cytotoxic.
[0101] The diseases or disorders treatable with the methods provided herein include, but
are not limited to, cancer, disorders associated with angiogenesis, pain including
Complex Regional Pain Syndrome ("CRPS"), Macular Degeneration ("MD") and related syndromes,
skin diseases, pulmonary disorders, asbestos-related disorders, parasitic diseases,
immunodeficiency disorders, CNS disorders, CNS injury, atherosclerosis and related
disorders, dysfunctional sleep and related disorders, hemoglobinopathy and related
disorders (
e.g., anemia), TNFα and other cytokines related disorders, and other various diseases
and disorders.
[0102] Examples of cancer and precancerous conditions include, but are not limited to, those
described in
U.S. patent nos. 6,281,230 and
5,635,517 to Muller et al., in various U.S. patent publications to Zeldis, including Publ. Nos.
2004/0220144A1, published November 4, 2004 (Treatment of Myelodysplastic Syndrome);
2004/0029832A1, published February 12, 2004 (Treatment of Various Types of Cancer); and
2004/0087546, published May 6, 2004 (Treatment of Myeloproliferative Diseases). Examples also include those described
in
PCT/US04/14004, filed May 5, 2004.
[0103] Examples of cancer include, but are not limited to, cancers of the skin, such as
melanoma; lymph node; breast; cervix; uterus; gastrointestinal tract; lung; ovary;
prostate; colon; rectum; mouth; brain; head and neck; throat; testes; thyroid; kidney;
pancreas; bone; spleen; liver; bladder; larynx; nasal passages; and AIDS-related cancers.
The compounds provided here are useful for treating cancers of the blood and bone
marrow, such as multiple myeloma and acute and chronic leukemias, for example, lymphoblastic,
myelogenous, lymphocytic, and myelocytic leukemias. The compounds provided herein
can be used for treating, preventing, or managing either primary or metastatic tumors.
[0104] Other examples of cancers include, but are not limited to, advanced malignancy, amyloidosis,
neuroblastoma, meningioma, hemangiopericytoma, multiple brain metastase, glioblastoma
multiforms, glioblastoma, brain stem glioma, poor prognosis malignant brain tumor,
malignant glioma, recurrent malignant glioma, anaplastic astrocytoma, anaplastic oligodendroglioma,
neuroendocrine tumor, rectal adenocarcinoma, Dukes C & D colorectal cancer, unresectable
colorectal carcinoma, metastatic hepatocellular carcinoma, Kaposi's sarcoma, karotype
acute myeloblastic leukemia, chronic lymphocytic leukemia (CLL), Hodgkin's lymphoma,
non-Hodgkin's lymphoma, cutaneous T-Cell lymphoma, cutaneous B-Cell lymphoma, diffuse
large B-Cell lymphoma, low grade follicular lymphoma, metastatic melanoma (localized
melanoma, including, but not limited to, ocular melanoma), malignant mesothelioma,
malignant pleural effusion mesothelioma syndrome, peritoneal carcinoma, papillary
serous carcinoma, gynecologic sarcoma, soft tissue sarcoma, scleroderma, cutaneous
vasculitis, Langerhans cell histiocytosis, leiomyosarcoma, fibrodysplasia ossificans
progressive, hormone refractory prostate cancer, resected high-risk soft tissue sarcoma,
unreseectable hepatocellular carcinoma, Waldenstrom's macroglobulinemia, smoldering
myeloma, indolent myeloma, fallopian tube cancer, androgen independent prostate cancer,
androgen dependent stage IV non-metastatic prostate cancer, hormone-insensitive prostate
cancer, chemotherapy-insensitive prostate cancer, papillary thyroid carcinoma, follicular
thyroid carcinoma, medullary thyroid carcinoma, and leiomyoma. In certain embodiments,
the cancer is metastatic. In certain embodiments, the cancer is refractory or resistance
to chemotherapy or radiation.
[0105] In another embodiment, provided herein are compounds and pharmaceutical compositions
of the invention for use in methods of treating, preventing or managing various forms
of leukemias such as chronic lymphocytic leukemia, chronic myelocytic leukemia, acute
lymphoblastic leukemia, acute myelogenous leukemia and acute myeloblastic leukemia,
including leukemias that are relapsed, refractory or resistant, as disclosed in U.S.
Publ. No.
2006/0030594, published February 9, 2006. The term "leukemia" refers malignant neoplasms of the blood-forming tissues. The
leukemia includes, but is not limited to, chronic lymphocytic leukemia, chronic myelocytic
leukemia, acute lymphoblastic leukemia, acute myelogenous leukemia and acute myeloblastic
leukemia. The leukemia can be relapsed, refractory or resistant to conventional therapy.
The term "relapsed" refers to a situation where patients who have had a remission
of leukemia after therapy have a return of leukemia cells in the marrow and a decrease
in normal blood cells. The term "refractory or resistant" refers to a circumstance
where patients, even after intensive treatment, have residual leukemia cells in their
marrow.
[0106] In yet another embodiment, provided herein are methods of treating, preventing or
managing various types of lymphomas, including Non-Hodgkin's lymphoma (NHL). The term
"lymphoma" refers a heterogenous group of neoplasms arising in the reticuloendothelial
and lymphatic systems. "NHL" refers to malignant monoclonal proliferation of lymphoid
cells in sites of the immune system, including lymph nodes, bone marrow, spleen, liver
and gastrointestinal tract. Examples of NHL include, but are not limited to, mantle
cell lymphoma, MCL, lymphocytic lymphoma of intermediate differentiation, intermediate
lymphocytic lymphoma, ILL, diffuse poorly differentiated lymphocytic lymphoma, PDL,
centrocytic lymphoma, diffuse small-cleaved cell lymphoma, DSCCL, follicular lymphoma,
and any type of the mantle cell lymphomas that can be seen under the microscope (nodular,
diffuse, blastic and mentle zone lymphoma).
[0107] Examples of diseases and disorders associated with, or characterized by, undesired
angiogenesis include, but are not limited to, inflammatory diseases, autoimmune diseases,
viral diseases, genetic diseases, allergic diseases, bacterial diseases, ocular neovascular
diseases, choroidal neovascular diseases, retina neovascular diseases, and rubeosis
(neovascularization of the angle). Specific examples of the diseases and disorders
associated with, or characterized by, undesired angiogenesis include, but are not
limited to, endometriosis, Crohn's disease, heart failure, advanced heart failure,
renal impairment, endotoxemia, toxic shock syndrome, osteoarthritis, retrovirus replication,
wasting, meningitis, silica-induced fibrosis, asbestos-induced fibrosis, veterinary
disorder, malignancy-associated hypercalcemia, stroke, circulatory shock, periodontitis,
gingivitis, macrocytic anemia, refractory anemia, and 5q-deletion syndrome.
[0108] Examples of pain include, but are not limited to those described in U.S. patent Publ.
No. 2005/0203142, published September 15, 2005. Specific types of pain include, but are not limited to, nociceptive pain, neuropathic
pain, mixed pain of nociceptive and neuropathic pain, visceral pain, migraine, headache,
and post-operative pain.
[0109] Examples of nociceptive pain include, but are not limited to, pain associated with
chemical or thermal burns, cuts of the skin, contusions of the skin, osteoarthritis,
rheumatoid arthritis, tendonitis, and myofascial pain.
[0110] Examples of neuropathic pain include, but are not limited to, CRPS type I, CRPS type
II, reflex sympathetic dystrophy (RSD), reflex neurovascular dystrophy, reflex dystrophy,
sympathetically maintained pain syndrome, causalgia, Sudeck atrophy of bone, algoneurodystrophy,
shoulder hand syndrome, post-traumatic dystrophy, trigeminal neuralgia, post herpetic
neuralgia, cancer related pain, phantom limb pain, fibromyalgia, chronic fatigue syndrome,
spinal cord injury pain, central post-stroke pain, radiculopathy, diabetic neuropathy,
post-stroke pain, luetic neuropathy, and other painful neuropathic conditions such
as those induced by drugs such as vincristine and velcade. As used herein, the terms
"complex regional pain syndrome," "CRPS" and "CRPS and related syndromes" mean a chronic
pain disorder characterized by one or more of the following: pain, whether spontaneous
or evoked, including allodynia (painful response to a stimulus that is not usually
painful) and hyperalgesia (exaggerated response to a stimulus that is usually only
mildly painful); pain that is disproportionate to the inciting event (
e.g., years of severe pain after an ankle sprain); regional pain that is not limited
to a single peripheral nerve distribution; and autonomic dysregulation (
e.g., edema, alteration in blood flow and hyperhidrosis) associated with trophic skin
changes (hair and nail growth abnormalities and cutaneous ulceration).
[0111] Examples of MD and related syndromes include, but are not limited to, those described
in U.S. patent Publ. No.
2004/0091455, published May 13, 2004. Specific examples include, but are not limited to, atrophic (dry) MD, exudative
(wet) MD, age-related maculopathy (ARM), choroidal neovascularisation (CNVM), retinal
pigment epithelium detachment (PED), and atrophy of retinal pigment epithelium (RPE).
[0112] Examples of skin diseases include, but are not limited to, those described in
U.S. Publ. No. 2005/0214328A1, published September 29, 2005. Specific examples include, but are not limited to,
keratoses and related symptoms, skin diseases or disorders characterized with overgrowths
of the epidermis, acne, and wrinkles. As used herein, the term "keratosis" refers
to any lesion on the epidermis marked by the presence of circumscribed overgrowths
of the horny layer, including but not limited to actinic keratosis, seborrheic keratosis,
keratoacanthoma, keratosis follicularis (Darier disease), inverted follicular keratosis,
palmoplantar keratoderma (PPK, keratosis palmaris et plantaris), keratosis pilaris,
and stucco keratosis. The term "actinic keratosis" also refers to senile keratosis,
keratosis senilis, verruca senilis, plana senilis, solar keratosis, keratoderma or
keratoma. The term "seborrheic keratosis" also refers to seborrheic wart, senile wart,
or basal cell papilloma. Keratosis is characterized by one or more of the following
symptoms: rough appearing, scaly, erythematous papules, plaques, spicules or nodules
on exposed surfaces (
e.g., face, hands, ears, neck, legs and thorax), excrescences of keratin referred to
as cutaneous horns, hyperkeratosis, telangiectasias, elastosis, pigmented lentigines,
acanthosis, parakeratosis, dyskeratoses, papillomatosis, hyperpigmentation of the
basal cells, cellular atypia, mitotic figures, abnormal cell-cell adhesion, dense
inflammatory infiltrates and small prevalence of squamous cell carcinomas.
[0113] Examples of skin diseases or disorders characterized with overgrowths of the epidermis
include, but are not limited to, any conditions, diseases or disorders marked by the
presence of overgrowths of the epidermis, including but not limited to, infections
associated with papilloma virus, arsenical keratoses, sign of Leser-Trélat, warty
dyskeratoma (WD), trichostasis spinulosa (TS), erythrokeratodermia variabilis (EKV),
ichthyosis fetalis (harlequin ichthyosis), knuckle pads, cutaneous melanoacanthoma,
porokeratosis, psoriasis, squamous cell carcinoma, confluent and reticulated papillomatosis
(CRP), acrochordons, cutaneous horn, cowden disease (multiple hamartoma syndrome),
dermatosis papulosa nigra (DPN), epidermal nevus syndrome (ENS), ichthyosis vulgaris,
molluscum contagiosum, prurigo nodularis, and acanthosis nigricans (AN).
[0114] Examples of pulmonary disorders include, but are not limited to, those described
in
U.S. Publ. No. 2005/0239842A1, published October 27, 2005. Specific examples include pulmonary hypertension and related disorders. Examples
of pulmonary hypertension and related disorders include, but are not limited to: primary
pulmonary hypertension (PPH); secondary pulmonary hypertension (SPH); familial PPH;
sporadic PPH; precapillary pulmonary hypertension; pulmonary arterial hypertension
(PAH); pulmonary artery hypertension; idiopathic pulmonary hypertension; thrombotic
pulmonary arteriopathy (TPA); plexogenic pulmonary arteriopathy; functional classes
I to IV pulmonary hypertension; and pulmonary hypertension associated with, related
to, or secondary to, left ventricular dysfunction, mitral valvular disease, constrictive
pericarditis, aortic stenosis, cardiomyopathy, mediastinal fibrosis, anomalous pulmonary
venous drainage, pulmonary venoocclusive disease, collagen vasular disease, congenital
heart disease, HIV virus infection, drugs and toxins such as fcnfluramincs, congenital
heart disease, pulmonary venous hypertension, chronic obstructive pulmonary disease,
interstitial lung disease, sleep-disordered breathing, alveolar hypoventilation disorder,
chronic exposure to high altitude, neonatal lung disease, alveolar-capillary dysplasia,
sickle cell disease, other coagulation disorder, chronic thromboemboli, connective
tissue disease, lupus including systemic and cutaneous lupus, schistosomiasis, sarcoidosis
or pulmonary capillary hemangiomatosis.
[0115] Examples of asbestos-related disorders include, but not limited to, those described
in
U.S. Publ. No. 2005/0100529, published May 12, 2005. Specific examples include, but are not limited to, mesothelioma, asbestosis, malignant
pleural effusion, benign exudative effusion, pleural plaques, pleural calcification,
diffuse pleural thickening, rounded atelectasis, fibrotic masses, and lung cancer.
[0116] Examples of parasitic diseases include, but are not limited to, those described in
U.S. Publ. No. 2006/0154880, published July 13, 2006. Parasitic diseases include diseases and disorders caused by human intracellular
parasites such as, but not limited to,
P. falcifarium, P. ovale, P. vivax, P. malariae, L. donovari, L. infantum, L. aethiopica,
L. major, L. tropica, L. mexicana, L. braziliensis, T. Gondii, B. microti, B. divergens,
B. coli, C. parvum, C. cayetanensis, E. histolytica, I. belli, S. mansonii, S. haematobium,
Trypanosoma ssp., Toxoplasma ssp., and
O.
volvulus. Other diseases and disorders caused by non-human intracellular parasites such as,
but not limited to,
Babesia bovis, Babesia canis, Banesia Gibsoni, Besnoitia darlingi, Cytauxzoon felis,
Eimeria ssp., Hammondia ssp., and
Theileria ssp., are also encompassed. Specific examples include, but are not limited to, malaria,
babesiosis, trypanosomiasis, leishmaniasis, toxoplasmosis, meningoencephalitis, keratitis,
amebiasis, giardiasis, cryptosporidiosis, isosporiasis, cyclosporiasis, microsporidiosis,
ascariasis, trichuriasis, ancylostomiasis, strongyloidiasis, toxocariasis, trichinosis,
lymphatic filariasis, onchocerciasis, filariasis, schistosomiasis, and dermatitis
caused by animal schistosomes.
[0117] Examples of immunodeficiency disorders include, but are not limited to, those described
in
U.S. Publ. No. 2006/0188475, published August 24, 2006. Specific examples include, but not limited to, adenosine deaminase deficiency, antibody
deficiency with normal or elevated Igs, ataxia-tenlangiectasia, bare lymphocyte syndrome,
common variable immunodeficiency, Ig deficiency with hyper-IgM, Ig heavy chain deletions,
IgA deficiency, immunodeficiency with thymoma, reticular dysgenesis, Nezelof syndrome,
selective IgG subclass deficiency, transient hypogammaglobulinemia of infancy, Wistcott-Aldrich
syndrome, X-linked agammaglobulinemia, X-linked severe combined immunodeficiency.
[0118] Examples of CNS disorders include, but are not limited to, those described in
U.S. Publ. No. 2005/0143344A1, published June 30, 2005. Specific examples include, but are not limited to, include, but are not limited
to, Amyotrophic Lateral Sclerosis, Alzheimer Disease, Parkinson Disease, Huntington's
Disease, Multiple Sclerosis other neuroimmunological disorders such as Tourette Syndrome,
delerium, or disturbances in consciousness that occur over a short period of time,
and amnestic disorder, or discreet memory impairments that occur in the absence of
other central nervous system impairments.
[0119] Examples of CNS injuries and related syndromes include, but are not limited to, those
described in
U.S. Publ. No. 2006/0122228, published June 8, 2006. Specific examples include, but are not limited to, CNS injury/damage and related
syndromes, include, but are not limited to, primary brain injury, secondary brain
injury, traumatic brain injury, focal brain injury, diffuse axonal injury, head injury,
concussion, post-concussion syndrome, cerebral contusion and laceration, subdural
hematoma, epidermal hematoma, post-traumatic epilepsy, chronic vegetative state, complete
SCI, incomplete SCI, acute SCI, subacute SCI, chronic SCI, central cord syndrome,
Brown-Sequard syndrome, anterior cord syndrome, conus medullaris syndrome, cauda equina
syndrome, neurogenic shock, spinal shock, altered level of consciousness, headache,
nausea, emesis, memory loss, dizziness, diplopia, blurred vision, emotional lability,
sleep disturbances, irritability, inability to concentrate, nervousness, behavioral
impairment, cognitive deficit, and seizure.
[0120] Other disease or disorders include, but not limited to, viral, genetic, allergic,
and autoimmune diseases. Specific examples include, but not limited to, HIV, hepatitis,
adult respiratory distress syndrome, bone resorption diseases, chronic pulmonary inflammatory
diseases, dermatitis, cystic fibrosis, septic shock, sepsis, endotoxic shock, hemodynamic
shock, sepsis syndrome, post ischemic reperfusion injury, meningitis, psoriasis, fibrotic
disease, cachexia, graft versus host disease, graft rejection, auto-immune disease,
rheumatoid spondylitis, Crohn's disease, ulcerative colitis, inflammatory-bowel disease,
multiple sclerosis, systemic lupus erythrematosus, ENL in leprosy, radiation damage,
cancer, asthma, or hyperoxic alveolar injury.
[0121] Examples of atherosclerosis and related conditions include, but are not limited to,
those disclosed in
U.S. Publ. No. 2002/0054899, published May 9, 2002. Specific examples include, but are not limited to, all forms of conditions involving
atherosclerosis, including restenosis after vascular intervention such as angioplasty,
stenting, atherectomy and grafting. All forms of vascular intervention are contemplated
by the disclosure, including diseases of the cardiovascular and renal system, such
as, but not limited to, renal angioplasty, percutaneous coronary intervention (PCI),
percutaneous transluminal coronary angioplasty (PTCA), carotid percutaneous transluminal
angioplasty (PTA), coronary by-pass grafting, angioplasty with stent implantation,
peripheral percutaneous transluminal intervention of the iliac, femoral or popliteal
arteries, and surgical intervention using impregnated artificial grafts. The following
chart provides a listing of the major systemic arteries that may be in need of treatment,
all of which are contemplated by the disclosure:
Artery |
Body Area Supplied |
Axillary |
Shoulder and axilla |
Brachial |
Upper arm |
Brachiocephalic |
Head, neck, and arm |
Celiac |
Divides into left gastric, splenic, and hepatic arteries |
Common carotid |
Neck |
Common iliac |
Divides into external and internal iliac arteries |
Coronary |
Heart |
Deep femoral |
Thigh |
Digital |
Fingers |
Dorsalis pedis |
Foot |
External carotid |
Neck and external head regions |
External iliac |
Femoral artery |
Femoral |
Thigh |
Gastric |
Stomach |
Hepatic |
Liver, gallbladder, pancreas, and duodenum |
Inferior mesenteric |
Descending colon, rectum, and pelvic wall |
Internal carotid |
Neck and internal head regions |
Internal iliac |
Rectum, urinary bladder, external genitalia, buttocks muscles, uterus and vagina |
Left gastric |
Esophagus and stomach |
Middle sacral |
Sacrum |
Ovarian |
Ovaries |
Palmar arch |
Hand |
Peroneal |
Calf |
Popliteal |
Knee |
Posterior tibial |
Calf |
Pulmonary |
Lungs |
Radial |
Forearm |
Renal |
Kidney |
Splenic |
Stomach, pancreas, and spleen |
Subclavian |
Shoulder |
Superior mesenteric |
Pancreas, small intestine, ascending and transverse colon |
Testicular |
Testes |
Ulnar |
Forearm |
[0122] Examples of dysfunctional sleep and related syndromes include, but are not limited
to, those disclosed in
U.S. Publ. No. 2005/0222209A1, published October 6, 2005. Specific examples include, but are not limited to, snoring, sleep apnea, insomnia,
narcolepsy, restless leg syndrome, sleep terrors, sleep walking sleep eating, and
dysfunctional sleep associated with chronic neurological or inflammatory conditions.
Chronic neurological or inflammatory conditions, include, but are not limited to,
Complex Regional Pain Syndrome, chronic low back pain, musculoskeletal pain, arthritis,
radiculopathy, pain associated with cancer, fibromyalgia, chronic fatigue syndrome,
visceral pain, bladder pain, chronic pancreatitis, neuropathies (diabetic, post-herpetic,
traumatic or inflammatory), and neurodegenerative disorders such as Parkinson's Disease,
Alzheimer's Disease, amyotrophic lateral sclerosis, multiple sclerosis, Huntington's
Disease, bradykinesia; muscle rigidity; parkinsonian tremor; parkinsonian gait; motion
freezing; depression; defective long-term memory, Rubinstein-Taybi syndrome (RTS);
dementia; postural instability; hypokinetic disorders; synuclein disorders; multiple
system atrophies; striatonigral degeneration; olivopontocerebellar atrophy; Shy-Drager
syndrome; motor neuron disease with parkinsonian features; Lewy body dementia; Tau
pathology disorders; progressive supranuclear palsy; corticobasal degeneration; frontotemporal
dementia; amyloid pathology disorders; mild cognitive impairment; Alzheimer disease
with parkinsonism; Wilson disease; Hallervorden-Spatz disease; Chediak-Hagashi disease;
SCA-3 spinocerebellar ataxia; X-linked dystonia parkinsonism; prion disease; hyperkinetic
disorders; chorea; ballismus; dystonia tremors; Amyotrophic Lateral Sclerosis (ALS);
CNS trauma and myoclonus.
[0123] Examples of hemoglobinopathy and related disorders include, but are not limited to,
those described in
U.S. Publ. No. 2005/0143420A1,
published June 30, 2005. Specific examples include, but are not limited to, hemoglobinopathy, sickle cell
anemia, and any other disorders related to the differentiation of CD34+ cells.
[0124] Examples of TNFα and other cytokines related disorders include, but are not limited
to, those described in
WO 98/03502 and
WO 98/54170. Specific examples include, but are not limited to: endotoxemia or toxic shock syndrome;
cachexia; adult respiratory distress syndrome; bone resorption diseases such as arthritis;
hypercalcemia; Graft versus Host Reaction; cerebral malaria; inflammation; tumor growth;
chronic pulmonary inflammatory diseases; reperfusion injury; myocardial infarction;
stroke; circulatory shock; rheumatoid arthritis; Crohn's disease; HIV infection and
AIDS; other disorders such as rheumatoid arthritis, rheumatoid spondylitis, osteoarthritis
and other arthritic conditions, septic shock, septis, endotoxic shock, graft versus
host disease, wasting, Crohn's disease, ulcerative colitis, multiple sclerosis, systemic
lupus erythromatosis, ENL in leprosy, HIV, AIDS, and opportunistic infections in AIDS;
cAMP related disorders such as septic shock, sepsis, endotoxic shock, hemodynamic
shock and sepsis syndrome, post ischemic reperfusion injury, malaria, mycobacterial
infection, meningitis, psoriasis, congestive heart failure, fibrotic disease, cachexia,
graft rejection, oncogenic or cancerous conditions, asthma, autoimmune disease, radiation
damages, and hyperoxic alveolar injury; viral infections, such as those caused by
the herpes viruses; viral conjunctivitis; or atopic dermatitis.
[0125] In certain embodiments, the use of compounds provided herein in various immunological
applications, in particular, as vaccine adjuvants, particularly anticancer vaccine
adjuvants, as disclosed in
U.S. Publ. No. 2007/0048327, published March 1, 2007, is also encompassed. This aspect of the disclosure also relates to the uses of compounds
provided in combination with vaccines to treat or prevent cancer or infectious diseases,
and other various uses of immunomodulatory compounds such as reduction or desensitization
of allergic reactions.
[0126] Depending on the condition, disorder, or disease to be treated and the subject's
condition, a compound provided herein may be administered by oral, parenteral (
e.g., intramuscular, intraperitoneal, intravenous, ICV, intracistemal injection or infusion,
subcutaneous injection, or implant), inhalation, nasal, vaginal, rectal, sublingual,
and/or topical (e.g., transdermal or local) routes of administration, and may be formulated
alone or together in suitable dosage unit with a pharmaceutically acceptable vehicle,
carrier, diluent, excipient, or a mixture thereof, appropriate for each route of administration.
[0127] The dose may be in the form of one, two, three, four, five, six, or more sub-doses
that are administered at appropriate intervals per day. The dose or sub-doses can
be administered in the form of dosage units containing from about 0.1 to about 1,000
mg, from about 0.1 to about 500 mg, or from 0.5 about to about 100 mg of active ingredient(s)
per dosage unit, and if the condition of the patient requires, the dose can, by way
of alternative, be administered as a continuous infusion.
[0128] In certain embodiments, an appropriate dosage level is about 0.01 to about 100 mg
per kg patient body weight per day (mg/kg per day), about 0.01 to about 50 mg/kg per
day, about 0.01 to about 25 mg/kg per day, or about 0.05 to about 10 mg/kg per day,
which may be administered in single or multiple doses. A suitable dosage level may
be about 0.01 to about 100 mg/kg per day, about 0.05 to about 50 mg/kg per day, or
about 0.1 to about 10 mg/kg per day. Within this range, the dosage may be about 0.01
to about 0.1, about 0.1 to about 1.0, about 1.0 to about 10, or about 10 to about
50 mg/kg per day.
[0129] For oral administration, the pharmaceutical compositions can be provided in the form
of tablets containing 1.0 to 1,000 mg of the active ingredient, particularly about
1, about 5, about 10, about 15, about 20, about 25, about 50, about 75, about 100,
about 150, about 200, about 250, about 300, about 400, about 500, about 600, about
750, about 800, about 900, and about 1,000 mg of the active ingredient for the symptomatic
adjustment of the dosage to the patient to be treated. The compositions may be administered
on a regimen of 1 to 4 times per day, including once, twice, three times, and four
times per day.
[0130] It will be understood, however, that the specific dose level and frequency of dosage
for any particular patient may be varied and will depend upon a variety of factors
including the activity of the specific compound employed, the metabolic stability
and length of action of that compound, the age, body weight, general health, sex,
diet, mode and time of administration, rate of excretion, drug combination, the severity
of the particular condition, and the host undergoing therapy.
4.4 SECOND ACTIVE AGENTS
[0131] The compounds provided herein may also be combined or used in combination with other
therapeutic agents useful in treating, preventing, and/or managing various diseases
or disorders for which the compounds provided herein are useful. It is believed that
certain combinations may work synergistically in the treatment of particular types
of diseases or disorders, and conditions and symptoms associated with such diseases
or disorders. A compound provided herein, including a single enantiomer, a mixture
of enantiomers, or a mixture of diastereomers thereof, or a pharmaceutically acceptable
salt, solvate, or prodrug thereof, can also work to alleviate adverse effects associated
with certain second active agents, and vice versa.
[0132] As used herein, the term "in combination" includes the use of more than one therapeutic
agents. However, the use of the term "in combination" does not restrict the order
or route in which therapeutic agents are administered to a subject with a condition,
disorder, or disorder. A first therapeutic agent (
e.g., a therapeutic agent such as a compound provided herein) can be administered prior
to (
e.g., 5 min, 15 min, 30 min, 45 min, 1 hr, 2 hrs, 4 hrs, 6 hrs, 12 hrs, 24 hrs, 48 hrs,
72 hrs, 96 hrs, 1 wk, 2 wks, 3 wks, 4 wks, 5 wks, 6 wks, 8 wks, or 12 wks before),
concomitantly with, or subsequent to
(e.g., 5 min, 15 min, 30 min, 45 min, 1 hr, 2 hrs, 4 hrs, 6 hrs, 12 hrs, 24 hrs, 48 hrs,
72 hrs, 96 hrs, 1 wk, 2 wks, 3 wks, 4 wks, 5 wks, 6 wks, 8 wks, or 12 wks after) the
administration of a second therapeutic agent to a subject to be treated.
[0133] The suitability of a particular route of administration employed for a particular
active agent will depend on the active agent itself (
e.g., whether it can be administered orally without decomposing prior to entering the blood
stream) and the disease being treated. In certain embodiments, the route of administration
for compounds provided herein is oral. In certain embodiments, the routes of administration
for the second active agents or ingredients provided herein are those as described
in
Physicians' Desk Reference, 1755-1760 (56th ed., 2002).
[0134] In one embodiment, the second active agent is administered intravenously or subcutaneously
and once or twice daily in an amount of from about 1 to about 1,000 mg, from about
5 to about 500 mg, from about 10 to about 350 mg, or from about 50 to about 200 mg.
The specific amount of the second active agent will depend on the specific agent used,
the type of disease being treated or managed, the severity and stage of disease, and
the amount(s) of compounds provided herein and any optional additional active agents
concurrently administered to the patient.
[0135] Also described herein is a method of reducing, treating and/or preventing adverse
or undesired effects associated with conventional therapy including, but not limited
to, surgery, chemotherapy, radiation therapy, hormonal therapy, biological therapy
and immunotherapy. Compounds provided herein and other active ingredients can be administered
to a patient prior to, during, or after the occurrence of the adverse effect associated
with conventional therapy.
[0136] When a compound provided herein is used contemporaneously with one or more therapeutic
agents, a pharmaceutical composition containing such other agents in addition to the
compound provided herein may be utilized, but is not required. Accordingly, the pharmaceutical
compositions provided herein include those that also contain one or more other therapeutic
agents, in addition to a compound provided herein.
[0137] One or more second active ingredients or agents can be used in the methods and compositions
provided herein. Second active agents can be large molecules (
e.g., proteins) or small molecules (
e.g., synthetic inorganic, organometallic, or organic molecules).
[0138] Examples of large molecule active agents include, but are not limited to, hematopoietic
growth factors, cytokines, and monoclonal and polyclonal antibodies. Specific examples
of the active agents are anti-CD40 monoclonal antibodies (such as, for example, SGN-40,
Herceptin, rituximab); histone deacetlyase inhibitors (such as, for example, SAHA
and LAQ 824); heat-shock protein-90 inhibitors (such as, for example, 17-AAG); insulin-like
growth factor-1 receptor kinase inhibitors; vascular endothelial growth factor receptor
kinase inhibitors (such as, for example, PTK787); insulin growth factor receptor inhibitors;
lysophosphatidic acid acyltransrerase inhibitors; IkB kinase inhibitors; p38MAPK inhibitors;
EGFR inhibitors (such as, for example, gefitinib and erlotinib HCL); HER-2 antibodies
(such as, for example, trastuzumab (Herceptin®) and pertuzumab (Omnitarg™)); VEGFR
antibodies (such as, for example, bevacizumab (Avastin™)); VEGFR inhibitors (such
as, for example, flk-1 specific kinase inhibitors, SU5416 and ptk787/zk222584); P13K
inhibitors (such as, for example, wortmannin); C-Met inhibitors (such as, for example,
PHA-665752); monoclonal antibodies (such as, for example, rituximab (Rituxan®), tositumomab
(Bexxar®), edrecolomab (Panorex®) and G250); and anti-TNF-α antibodies. Examples of
small molecule active agents include, but are not limited to, small molecule anti-cancer
agents and antibiotics (
e.g., clarithromycin).
[0139] Specific second active compounds that can be combined with compounds provided herein
vary depending on the specific indication to be treated, prevented or managed.
[0140] For instance, for the treatment, prevention or management of cancer, second active
agents include, but are not limited to: semaxanib; cyclosporin; etanercept; doxycycline;
bortezomib; acivicin; aclarubicin; acodazole hydrochloride; acronine; adozelesin;
aldesleukin; altretamine; ambomycin; ametantrone acetate; amsacrine; anastrozole;
anthramycin; asparaginase; asperlin; azacitidine; azetepa; azotomycin; batimastat;
benzodepa; bicalutamide; bisantrene hydrochloride; bisnafide dimesylate; bizelesin;
bleomycin sulfate; brequinar sodium; bropirimine; busulfan; cactinomycin; calusterone;
caracemide; carbetimer; carboplatin; carmustine; carubicin hydrochloride; carzelesin;
cedefingol; celecoxib; chlorambucil; cirolemycin; cisplatin; cladribine; crisnatol
mesylate; cyclophosphamide; cytarabine; dacarbazine; dactinomycin; daunorubicin hydrochloride;
decitabine; dexormaplatin; dezaguanine; dezaguanine mesylate; diaziquone; docetaxel;
doxorubicin; doxorubicin hydrochloride; droloxifene; droloxifene citrate; dromostanolone
propionate; duazomycin; edatrexate; eflornithine hydrochloride; elsamitrucin; enloplatin;
enpromate; epipropidine; epirubicin hydrochloride; erbulozole; esorubicin hydrochloride;
estramustine; estramustine phosphate sodium; etanidazole; etoposide; etoposide phosphate;
etoprine; fadrozole hydrochloride; fazarabine; fenretinide; floxuridine; fludarabine
phosphate; fluorouracil; flurocitabine; fosquidone; fostriecin sodium; gemcitabine;
gemcitabine hydrochloride; hydroxyurea; idarubicin hydrochloride; ifosfamide; ilmofosine;
iproplatin; irinotecan; irinotecan hydrochloride; lanreotide acetate; letrozole; leuprolide
acetate; liarozole hydrochloride; lometrexol sodium; lomustine; losoxantrone hydrochloride;
masoprocol; maytansine; mechlorethamine hydrochloride; megestrol acetate; melengestrol
acetate; melphalan; menogaril; mercaptopurine; methotrexate; methotrexate sodium;
metoprine; meturedepa; mitindomide; mitocarcin; mitocromin; mitogillin; mitomalcin;
mitomycin; mitosper; mitotane; mitoxantrone hydrochloride; mycophenolic acid; nocodazole;
nogalamycin; ormaplatin; oxisuran; paclitaxel; pegaspargase; peliomycin; pentamustine;
peplomycin sulfate; perfosfamide; pipobroman; piposulfan; piroxantrone hydrochloride;
plicamycin; plomestane; porfimer sodium; porfiromycin; prednimustine; procarbazine
hydrochloride; puromycin; puromycin hydrochloride; pyrazofurin; riboprine; safingol;
safingol hydrochloride; semustine; simtrazene; sparfosate sodium; sparsomycin; spirogermanium
hydrochloride; spiromustine; spiroplatin; streptonigrin; streptozocin; sulofenur;
talisomycin; tecogalan sodium; taxotere; tegafur; teloxantrone hydrochloride; temoporfin;
teniposide; teroxirone; testolactone; thiamiprine; thioguanine; thiotepa; tiazofurin;
tirapazamine; toremifene citrate; trestolone acetate; triciribine phosphate; trimetrexate;
trimetrexate glucuronate; triptorelin; tubulozole hydrochloride; uracil mustard; uredepa;
vapreotide; verteporfin; vinblastine sulfate; vincristine sulfate; vindesine; vindesine
sulfate; vinepidine sulfate; vinglycinate sulfate; vinleurosine sulfate; vinorelbine
tartrate; vinrosidine sulfate; vinzolidine sulfate; vorozole; zeniplatin; zinostatin;
and zorubicin hydrochloride.
[0141] Other second agents include, but are not limited to: 20-epi-1,25 dihydroxyvitamin
D3; 5-ethynyluracil; abiraterone; aclarubicin; acylfulvene; adecypenol; adozelesin;
aldesleukin; ALL-TK antagonists; altretamine; ambamustine; amidox; amifostine; aminolevulinic
acid; amrubicin; amsacrine; anagrelide; anastrozole; andrographolide; angiogenesis
inhibitors; antagonist D; antagonist G; antarelix; anti-dorsalizing morphogenetic
protein-1; antiandrogen, prostatic carcinoma; antiestrogen; antineoplaston; antisense
oligonucleotides; aphidicolin glycinate; apoptosis gene modulators; apoptosis regulators;
apurinic acid; ara-CDP-DL-PTBA; arginine deaminase; asulacrine; atamestane; atrimustine;
axinastatin 1; axinastatin 2; axinastatin 3; azasetron; azatoxin; azatyrosine; baccatin
III derivatives; balanol; batimastat; BCR/ABL antagonists; benzochlorins; benzoylstaurosporine;
beta lactam derivatives; beta-alethine; betaclamycin B; betulinic acid; bFGF inhibitor;
bicalutamide; bisantrene; bisaziridinylspermine; bisnafide; bistratene A; bizelesin;
breflate; bropirimine; budotitane; buthionine sulfoximine; calcipotriol; calphostin
C; camptothecin derivatives; capecitabine; carboxamide-amino-triazole; carboxyamidotriazole;
CaRest M3; CARN 700; cartilage derived inhibitor; carzelesin; casein kinase inhibitors
(ICOS); castanospermine; cecropin B; cetrorelix; chlorlns; chloroquinoxaline sulfonamide;
cicaprost; cis-porphyrin; cladribine; clomifene analogues; clotrimazole; collismycin
A; collismycin B; combretastatin A4; combretastatin analogue; conagenin; crambescidin
816; crisnatol; cryptophycin 8; cryptophycin A derivatives; curacin A; cyclopentanthraquinones;
cycloplatam; cypemycin; cytarabine ocfosfate; cytolytic factor; cytostatin; dacliximab;
decitabine; dehydrodidemnin B; deslorelin; dexamethasone; dexifosfamide; dexrazoxane;
dexverapamil; diaziquone; didemnin B; didox; diethylnorspermine; dihydro-5-azacytidine;
dihydrotaxol; dioxamycin; diphenyl spiromustine; docetaxel; docosanol; dolasetron;
doxifluridine; doxorubicin; droloxifene; dronabinol; duocarmycin SA; ebselen; ecomustine;
edelfosine; edrecolomab; eflornithine; elemene; emitefur; epirubicin; epristeride;
estramustine analogue; estrogen agonists; estrogen antagonists; etanidazole; etoposide
phosphate; exemestane; fadrozole; fazarabine; fenretinide; filgrastim; finasteride;
flavopiridol; flezelastine; fluasterone; fludarabine; fluorodaunorunicin hydrochloride;
forfenimex; formestane; fostriecin; fotemustine; gadolinium texaphyrin; gallium nitrate;
galocitabine; ganirelix; gelatinase inhibitors; gemcitabine; glutathione inhibitors;
hepsulfam; heregulin; hexamethylene bisacetamide; hypericin; ibandronic acid; idarubicin;
idoxifene; idramantone; ilmofosine; ilomastat; imatinib (Gleevec®), imiquimod; immunostimulant
peptides; insulin-like growth factor-1 receptor inhibitor; interferon agonists; interferons;
interleukins; iobenguane; iododoxorubicin; ipomeanol, 4-; iroplact; irsogladine; isobengazole;
isohomohalicondrin B; itasetron; jasplakinolide; kahalalide F; lamellarin-N triacetate;
lanreotide; leinamycin; lenograstim; lentinan sulfate; leptolstatin; letrozole; leukemia
inhibiting factor; leukocyte alpha interferon; leuprolide+estrogen+progesterone; leuprorelin;
levamisole; liarozole; linear polyamine analogue; lipophilic disaccharide peptide;
lipophilic platinum compounds; lissoclinamide 7; lobaplatin; lombricine; lometrexol;
lonidamine; losoxantrone; loxoribine; lurtotecan; lutetium texaphyrin; lysofylline;
lytic peptides; maitansine; mannostatin A; marimastat; masoprocol; maspin; matrilysin
inhibitors; matrix metalloproteinase inhibitors; menogaril; merbarone; meterelin;
methioninase; metoclopramide; MIF inhibitor; mifepristone; miltefosine; mirimostim;
mitoguazone; mitolactol; mitomycin analogues; mitonafide; mitotoxin fibroblast growth
factor-saporin; mitoxantrone; mofarotene; molgramostim; Erbitux, human chorionic gonadotrophin;
monophosphoryl lipid A+myobacterium cell wall sk; mopidamol; mustard anticancer agent;
mycaperoxide B; mycobacterial cell wall extract; myriaporone; N-acetyldinaline; N-substituted
benzamides; nafarelin; nagrestip; naloxone+pentazocine; napavin; naphterpin; nartograstim;
nedaplatin; nemorubicin; neridronic acid; nilutamide; nisamycin; nitric oxide modulators;
nitroxide antioxidant; nitrullyn; oblimersen (Genasense®); O6-benzylguanine; octreotide;
okicenone; oligonucleotides; onapristone; ondansetron; ondansetron; oracin; oral cytokine
inducer; ormaplatin; osaterone; oxaliplatin; oxaunomycin; paclitaxel; paclitaxel analogues;
paclitaxel derivatives; palauamine; palmitoylrhizoxin; pamidronic acid; panaxytriol;
panomifene; parabactin; pazelliptine; pegaspargase; peldesine; pentosan polysulfate
sodium; pentostatin; pentrozole; perflubron; perfosfamide; perillyl alcohol; phenazinomycin;
phenylacetate; phosphatase inhibitors; picibanil; pilocarpine hydrochloride; pirarubicin;
piritrexim; placetin A; placetin B; plasminogen activator inhibitor; platinum complex;
platinum compounds; platinum-triamine complex; porfimer sodium; porfiromycin; prednisone;
propyl bis-acridone; prostaglandin J2; proteasome inhibitors; protein A-based immune
modulator; protein kinase C inhibitor; protein kinase C inhibitors, microalgal; protein
tyrosine phosphatase inhibitors; purine nucleoside phosphorylase inhibitors; purpurins;
pyrazoloacridine; pyridoxylated hemoglobin polyoxyethylene conjugate; raf antagonists;
raltitrexed; ramosetron; ras farnesyl protein transferase inhibitors; ras inhibitors;
ras-GAP inhibitor; retelliptine demethylated; rhenium Re 186 etidronate; rhizoxin;
ribozymes; RII retinamide; rohitukine; romurtide; roquinimex; rubiginone B1; ruboxyl;
safingol; saintopin; SarCNU; sarcophytol A; sargramostim; Sdi 1 mimetics; semustine;
senescence derived inhibitor 1; sense oligonucleotides; signal transduction inhibitors;
sizofiran; sobuzoxane; sodium borocaptate; sodium phenylacetate; solverol; somatomedin
binding protein; sonermin; sparfosic acid; spicamycin D; spiromustine; splenopentin;
spongistatin 1; squalamine; stipiamide; stromelysin inhibitors; sulfinosine; superactive
vasoactive intestinal peptide antagonist; suradista; suramin; swainsonine; tallimustine;
tamoxifen methiodide; tauromustine; tazarotene; tecogalan sodium; tegafur; tellurapyrylium;
telomerase inhibitors; temoporfin; teniposide; tetrachlorodecaoxide; tetrazomine;
thaliblastine; thiocoraline; thrombopoietin; thrombopoietin mimetic; thymalfasin;
thymopoietin receptor agonist; thymotrinan; thyroid stimulating hormone; tin ethyl
etiopurpurin; tirapazamine; titanocene bichloride; topsentin; toremifene; translation
inhibitors; tretinoin; triacetyluridine; triciribine; trimetrexate; triptorelin; tropisetron;
turosteride; tyrosine kinase inhibitors; tyrphostins; UBC inhibitors; ubenimex; urogenital
sinus-derived growth inhibitory factor; urokinase receptor antagonists; vapreotide;
variolin B; velaresol; veramine; verdins; verteporfin; vinorelbine; vinxaltine; vitaxin;
vorozole; zanoterone; zeniplatin; zilascorb; and zinostatin stimalamer.
[0142] Specific second active agents include, but are not limited to, 2-methoxyestradiol,
telomestatin, inducers of apoptosis in mutiple myeloma cells (such as, for example,
TRAIL), statins, semaxanib, cyclosporin, etanercept, doxycycline, bortezomib, oblimersen
(Genasense®), remicade, docetaxel, celecoxib, melphalan, dexamethasone (Decadron®),
steroids, gemcitabine, cisplatinum, temozolomide, etoposide, cyclophosphamide, temodar,
carboplatin, procarbazine, gliadel, tamoxifen, topotecan, methotrexate, Arisa®, taxol,
taxotere, fluorouracil, leucovorin, irinotecan, xeloda, CPT-11, interferon alpha,
pegylated interferon alpha (
e.g., PEG INTRON-A), capecitabine, cisplatin, thiotepa, fludarabine, carboplatin, liposomal
daunorubicin, cytarabine, doxetaxol, pacilitaxel, vinblastine, IL-2, GM-CSF, dacarbazine,
vinorelbine, zoledronic acid, palmitronate, biaxin, busulphan, prednisone, bisphosphonate,
arsenic trioxide, vincristine, doxorubicin (Doxil®), paclitaxel, ganciclovir, adriamycin,
estramustine sodium phosphate (Emcyt®), sulindac, and etoposide.
[0143] Similarly, examples of specific second agents according to the indications to be
treated, prevented, or managed can be found in the following references:
U.S. Pat. Nos. 6,281,230 and
5,635,517;
U.S. Appl. Nos. 10/411,649,
10/483,213,
10/411,656,
10/693,794,
10/699,154, and
10/981,189; and
U.S. Prov. Appl. Nos. 60/554,923,
60/565,172,
60/626,975,
60/630,599,
60/631,870, and
60/533,862.
[0144] Examples of second active agents that may be used for the treatment, prevention and/or
management of pain include, but are not limited to, conventional therapeutics used
to treat or prevent pain such as antidepressants, anticonvulsants, antihypertensives,
anxiolytics, calcium channel blockers, muscle relaxants, non-narcotic analgesics,
opioid analgesics, anti-inflammatories, cox-2 inhibitors, immunomodulatory agents,
alpha-adrenergic receptor agonists or antagonists, immunosuppressive agents, corticosteroids,
hyperbaric oxygen, ketamine, other anesthetic agents, NMDA antagonists, and other
therapeutics found, for example, in the
Physician's Desk Reference 2003. Specific examples include, but are not limited to, salicylic acid acetate (Aspirin®),
celecoxib (Celebrex®), Enbrel®, ketamine, gabapentin (Neurontin®), phenytoin (Dilantin®),
carbamazepine (Tegretol®), oxcarbazepine (Trileptal®), valproic acid (Depakene®),
morphine sulfate, hydromorphone, prednisone, griseofulvin, penthonium, alendronate,
dyphenhydramide, guanethidine, ketorolac (Acular®), thyrocalcitonin, dimethylsulfoxide
(DMSO), clonidine (Catapress®), bretylium, ketanserin, reserpine, droperidol, atropine,
phentolamine, bupivacaine, lidocaine, acetaminophen, nortriptyline (Pamelor®), amitriptyline
(Elavil®), imipramine (Tofranil®), doxepin (Sinequan®), clomipramine (Anafranil®),
fluoxetine (Prozac®), sertraline (Zoloft®), nefazodone (Serzone®), venlafaxine (Effexor®),
trazodone (Desyrel®), bupropion (Wellbutrin®), mexiletine, nifedipine, propranolol,
tramadol, lamotrigine, ziconotide, ketamine, dextromethorphan, benzodiazepines, baclofen,
tizanidine and phenoxybenzamine.
[0145] Examples of second active agents that may be used for the treatment, prevention and/or
management of MD and related syndromes include, but are not limited to, a steroid,
a light sensitizer, an integrin, an antioxidant, an interferon, a xanthine derivative,
a growth hormone, a neutrotrophic factor, a regulator of neovascularization, an anti-VEGF
antibody, a prostaglandin, an antibiotic, a phytoestrogen, an anti-inflammatory compound
or an antiangiogenesis compound, or a combination thereof. Specific examples include,
but are not limited to, verteporfin, purlytin, an angiostatic steroid, rhuFab, interferon-2α,
pentoxifylline, tin etiopurpurin, motexafin lutetium, 9-fluoro-11,21-dihydroxy-16,
17-1-methylethylidine-bis(oxy)pregna-1,4-diene-3,20-dione, latanoprost (see
U.S. Patent No. 6,225,348), tetracycline and its derivatives, rifamycin and its derivatives, macrolides, metronidazole
(
U.S. Patent Nos. 6,218,369 and
6,015,803), genistein, genistin, 6'-O-Mal genistin, 6'-O-Ac genistin, daidzein, daidzin, 6'-O-Mal
daidzin, 6'-O-Ac daidzin, glycitein, glycitin, 6'-O-Mal glycitin, biochanin A, formononetin
(
U.S. Patent No. 6,001,368), triamcinolone acetomide, dexamethasone (
U.S. Patent No. 5,770,589), thalidomide, glutathione (
U.S. Patent No. 5,632,984), basic fibroblast growth factor (bFGF), transforming growth factor b (TGF-b), brain-derived
neurotrophic factor (BDNF), plasminogen activator factor type 2 (PAI-2), EYE101 (Eyetech
Pharmaceuticals), LY333531 (Eli Lilly), Miravant, and RETISERT implant (Bausch & Lomb).
[0146] Examples of second active agents that may be used for the treatment, prevention and/or
management of skin diseases include, but are not limited to, keratolytics, retinoids,
α-hydroxy acids, antibiotics, collagen, botulinum toxin, interferon, and immunomodulatory
agents. Specific examples include, but are not limited to, 5-fluorouracil, masoprocol,
trichloroacetic acid, salicylic acid, lactic acid, ammonium lactate, urea, tretinoin,
isotretinoin, antibiotics, collagen, botulinum toxin, interferon, corticosteroid,
transretinoic acid and collagens such as human placental collagen, animal placental
collagen, Dermalogen, AlloDerm, Fascia, Cymetra, Autologen, Zyderm, Zyplast, Resoplast,
and Isolagen.
[0147] Examples of second active agents that may be used for the treatment, prevention and/or
management of pulmonary hepertension and related disorders include, but are not limited
to, anticoagulants, diuretics, cardiac glycosides, calcium channel blockers, vasodilators,
prostacyclin analogues, endothelin antagonists, phosphodiesterase inhibitors (e.g.,
PDE V inhibitors), endopeptidase inhibitors, lipid lowering agents, thromboxane inhibitors,
and other therapeutics known to reduce pulmonary artery pressure. Specific examples
include, but are not limited to, warfarin (Coumadin®), a diuretic, a cardiac glycoside,
digoxin-oxygen, diltiazem, nifedipine, a vasodilator such as prostacyclin (
e.g., prostaglandin I2 (PGI2), epoprostenol (EPO, Floran®), treprostinil (Remodulin®),
nitric oxide (NO), bosentan (Tracleer®), amlodipine, epoprostenol (Floran®), treprostinil
(Remodulin®), prostacyclin, tadalafil (Cialis®), simvastatin (Zocor®), omapatrilat
(Vanlev®), irbesartan (Avapro®), pravastatin (Pravachol®), digoxin, L-arginine, iloprost,
betaprost, and sildcnafil (Viagra®).
[0148] Examples of second active agents that may be used for the treatment, prevention and/or
management of asbestos-related disorders include, but are not limited to, anthracycline,
platinum, alkylating agent, oblimersen (Genasense®), cisplatinum, cyclophosphamide,
temodar, carboplatin, procarbazine, gliadel, tamoxifen, topotecan, methotrexate, taxotere,
irinotecan, capecitabine, cisplatin, thiotepa, fludarabine, carboplatin, liposomal
daunorubicin, cytarabine, doxetaxol, pacilitaxel, vinblastine, IL-2, GM-CSF, dacarbazine,
vinorelbine, zoledronic acid, palmitronate, biaxin, busulphan, prednisone, bisphosphonate,
arsenic trioxide, vincristine, doxorubicin (Doxil®), paclitaxel, ganciclovir, adriamycin,
bleomycin, hyaluronidase, mitomycin C, mepacrine, thiotepa, tetracycline and gemcitabine.
[0149] Examples of second active agents that may be used for the treatment, prevention and/or
management of parasitic diseases include, but are not limited to, chloroquine, quinine,
quinidine, pyrimethamine, sulfadiazine, doxycycline, clindamycin, mefloquine, halofantrine,
primaquine, hydroxychloroquine, proguanil, atovaquone, azithromycin, suramin, pentamidine,
melarsoprol, nifurtimox, benznidazole, amphotericin B, pentavalent antimony compounds
(
e.g., sodium stiboglucuronate), interfereon gamma, itraconazole, a combination of dead
promastigotes and BCG, leucovorin, corticosteroids, sulfonamide, spiramycin, IgG (serology),
trimethoprim, and sulfamethoxazole.
[0150] Examples of second active agents that may be used for the treatment, prevention and/or
management of immunodeficiency disorders include, but are not limited to: antibiotics
(therapeutic or prophylactic) such as, but not limited to, ampicillin, clarithromycin,
tetracycline, penicillin, cephalosporins, streptomycin, kanamycin, and erythromycin;
antivirals such as, but not limited to, amantadine, rimantadine, acyclovir, and ribavirin;
immunoglobulin; plasma; immunologic enhancing drugs such as, but not limited to, levami
sole and isoprinosine; biologics such as, but not limited to, gammaglobulin, transfer
factor, interleukins, and interferons; hormones such as, but not limited to, thymic;
and other immunologic agents such as, but not limited to, B cell stimulators (
e.g., BAFF/BlyS), cytokines (e.g., IL-2, IL-4, and IL-5), growth factors (
e.g., TGF-α), antibodies (
e.g., anti-CD40 and IgM), oligonucleotides containing unmethylated CpG motifs, and vaccines
(
e.g., viral and tumor peptide vaccines).
[0151] Examples of second active agents that may be used for the treatment, prevention and/or
management of CNS disorders include, but are not limited to: a dopamine agonist or
antagonist, such as, but not limited to, Levodopa, L-DOPA, cocaine, α-methyl-tyrosine,
reserpine, tetrabenazine, benzotropine, pargyline, fenodolpam mesylate, cabergoline,
pramipexole dihydrochloride, ropinorole, amantadine hydrochloride, selegiline hydrochloride,
carbidopa, pergolide mesylate, Sinemet CR, and Symmetrel; a MAO inhibitor, such as,
but not limited to, iproniazid, clorgyline, phenelzine and isocarboxazid; a COMT inhibitor,
such as, but not limited to, tolcapone and entacapone; a cholinesterase inhibitor,
such as, but not limited to, physostigmine saliclate, physostigmine sulfate, physostigmine
bromide, meostigmine bromide, neostigmine methylsulfate, ambenonim chloride, edrophonium
chloride, tacrine, pralidoxime chloride, obidoxime chloride, trimedoxime bromide,
diacetyl monoxim, endrophonium, pyridostigmine, and demecarium; an anti-inflammatory
agent, such as, but not limited to, naproxen sodium, diclofenac sodium, diclofenac
potassium, celecoxib, sulindac, oxaprozin, diflunisal, etodolac, meloxicam, ibuprofen,
ketoprofen, nabumetone, refecoxib, methotrexate, leflunomide, sulfasalazine, gold
salts, Rho-D Immune Globulin, mycophenylate mofetil, cyclosporine, azathioprine, tacrolimus,
basiliximab, daclizumab, salicylic acid, acetylsalicylic acid, methyl salicylate,
diflunisal, salsalate, olsalazine, sulfasalazine, acetaminophen, indomethacin, sulindac,
mefenamic acid, meclofenamate sodium, tolmetin, ketorolac, dichlofenac, flurbinprofen,
oxaprozin, piroxicam, meloxicam, ampiroxicam, droxicam, pivoxicam, tenoxicam, phenylbutazone,
oxyphenbutazone, antipyrine, aminopyrine, apazone, zileuton, aurothioglucose, gold
sodium thiomalate, auranofin, methotrexate, colchicine, allopurinol, probenecid, sulfinpyrazone
and benzbromarone or betamethasone and other glucocorticoids; and an antiemetic agent,
such as, but not limited to, metoclopromide, domperidone, prochlorperazine, promethazine,
chlorpromazine, trimethobenzamide, ondansetron, granisetron, hydroxyzine, acetylleucine
monoethanolamine, alizapride, azasetron, benzquinamide, bietanautine, bromopride,
buclizine, clebopride, cyclizine, dimenhydrinate, diphenidol, dolasetron, meclizine,
methallatal, metopimazine, nabilone, oxyperndyl, pipamazine, scopolamine, sulpiride,
tetrahydrocannabinol, thiethylperazine, thioproperazine, tropisetron, and a mixture
thereof.
[0152] Examples of second active agents that may be used for the treatment, prevention and/or
management of CNS injuries and related syndromes include, but are not limited to,
immunomodulatory agents, immunosuppressive agents, antihypertensives, anticonvulsants,
fibrinolytic agents, antiplatelet agents, antipsychotics, antidepressants, benzodiazepines,
buspirone, amantadine, and other known or conventional agents used in patients with
CNS injury/damage and related syndromes. Specific examples include, but are not limited
to: steroids (
e.g., glucocorticoids, such as, but not limited to, methylprednisolone, dexamethasone and
betamethasone); an anti-inflammatory agent, including, but not limited to, naproxen
sodium, diclofenac sodium, diclofenac potassium, celecoxib, sulindac, oxaprozin, diflunisal,
etodolac, meloxicam, ibuprofen, ketoprofen, nabumetone, refecoxib, methotrexate, leflunomide,
sulfasalazine, gold salts, RHo-D Immune Globulin, mycophenylate mofetil, cyclosporine,
azathioprine, tacrolimus, basiliximab, daclizumab, salicylic acid, acetylsalicylic
acid, methyl salicylate, diflunisal, salsalate, olsalazine, sulfasalazine, acetaminophen,
indomethacin, sulindac, mefenamic acid, meclofenamate sodium, tolmetin, ketorolac,
dichlofenac, flurbinprofen, oxaprozin, piroxicam, meloxicam, ampiroxicam, droxicam,
pivoxicam, tenoxicam, phenylbutazone, oxyphenbutazone, antipyrine, aminopyrine, apazone,
zileuton, aurothioglucose, gold sodium thiomalate, auranofin, methotrexate, colchicine,
allopurinol, probenecid, sulfinpyrazone and benzbromarone; a cAMP analog including,
but not limited to, db-cAMP; an agent comprising a methylphenidate drug, which comprises
1-threo-methylphenidate, d-threo-methylphenidate, dl-threo-methylphenidate, 1-erythro-methylphenidate,
d-erythro-methylphenidate, dl-erythro-methylphenidate, and a mixture thereof; and
a diuretic agent such as, but not limited to, mannitol, furosemide, glycerol, and
urea.
[0153] Examples of second active agent that may be used for the treatment, prevention and/or
management of dysfunctional sleep and related syndromes include, but are not limited
to, a tricyclic antidepressant agent, a selective serotonin reuptake inhibitor, an
antiepileptic agent (gabapentin, pregabalin, carbamazepine, oxcarbazepine, levitiracetam,
topiramate), an antiaryhthmic agent, a sodium channel blocking agent, a selective
inflammatory mediator inhibitor, an opioid agent, a second immunomodulatory compound,
a combination agent, and other known or conventional agents used in sleep therapy.
Specific examples include, but are not limited to, Neurontin, oxycontin, morphine,
topiramate, amitryptiline, nortryptiline, carbamazepine, Levodopa, L-DOPA, cocaine,
α-methyl-tyrosine, reserpine, tetrabenazine, benzotropine, pargyline, fenodolpam mesylate,
cabergoline, pramipexole dihydrochloride, ropinorole, amantadine hydrochloride, selegiline
hydrochloride, carbidopa, pergolide mesylate, Sinemet CR, Symmetrel, iproniazid, clorgyline,
phenelzine, isocarboxazid, tolcapone, entacapone, physostigmine saliclate, physostigmine
sulfate, physostigmine bromide, meostigmine bromide, neostigmine methylsulfate, ambenonim
chloride, edrophonium chloride, tacrine, pralidoxime chloride, obidoxime chloride,
trimedoxime bromide, diacetyl monoxim, endrophonium, pyridostigmine, demecarium, naproxen
sodium, diclofenac sodium, diclofenac potassium, celecoxib, sulindac, oxaprozin, diflunisal,
etodolac, meloxicam, ibuprofen, ketoprofen, nabumetone, refecoxib, methotrexate, leflunomide,
sulfasalazine, gold salts, RHo-D Immune Globulin, mycophenylate mofetil, cyclosporine,
azathioprine, tacrolimus, basiliximab, daclizumab, salicylic acid, acetylsalicylic
acid, methyl salicylate, diflunisal, salsalate, olsalazine, sulfasalazine, acetaminophen,
indomethacin, sulindac, mefenamic acid, meclofenamate sodium, tolmetin, ketorolac,
dichlofenac, flurbinprofen, oxaprozin, piroxicam, meloxicam, ampiroxicam, droxicam,
pivoxicam, tenoxicam, phenylbutazone, oxyphenbutazone, antipyrine, aminopyrine, apazone,
zileuton, aurothioglucose, gold sodium thiomalate, auranofin, methotrexate, colchicine,
allopurinol, probenecid, sulfinpyrazone, benzbromarone, betamethasone and other glucocorticoids,
metoclopromide, domperidone, prochlorperazine, promethazine, chlorpromazine, trimethobenzamide,
ondansetron, granisetron, hydroxyzine, acetylleucine monoethanolamine, alizapride,
azasetron, benzquinamide, bietanautine, bromopride, buclizine, clebopride, cyclizine,
dimenhydrinate, diphenidol, dolasctron, meclizine, mcthallatal, mctopimazinc, nabilonc,
oxyperndyl, pipamazine, scopolamine, sulpiride, tetrahydrocannabinol, thiethylperazine,
thioproperazine, tropisetron, and a mixture thereof.
[0154] Examples of second active agents that may be used for the treatment, prevention and/or
management of hemoglobinopathy and related disorders include, but are not limited
to: interleukins, such as IL-2 (including recombinant IL-II ("rIL2") and canarypox
IL-2), IL-10, IL-12, and IL-18; interferons, such as interferon alfa-2a, interferon
alfa-2b, interferon alfa-nl, interferon alfa-n3, interferon beta-I a, and interferon
gamma-I b; and G-CSF; hydroxyurea; butyrates or butyrate derivatives; nitrous oxide;
HEMOXIN™ (NIPRISAN™;
see United States Patent No.
5,800,819); Gardos channel antagonists such as clotrimazole and triaryl methane derivatives;
Deferoxamine; protein C; and transfusions of blood, or of a blood substitute such
as Hemospan™ or Hemospan™ PS (Sangart).
[0155] The dosages given will depend on absorption, inactivation and excretion rates of
the drug as well as other factors known to those of skill in the art. It is to be
noted that dosage values will also vary with the severity of the condition to be alleviated.
It is to be further understood that for any particular subject, specific dosage regimens
and schedules should be adjusted over time according to the individual need and the
professional judgment of the person administering or supervising the administration
of the compositions.
[0156] The weight ratio of a compound provided herein to the second active ingredient depends
upon the effective dose of each ingredient. Generally, an effective dose of each will
be used. Thus, for example, when a compound provided herein is combined with a PPAR
agonist the weight ratio of the compound provided herein to the PPAR agonist will
generally range from about 1000:1 to about 1:1000 or about 200:1 to about 1:200. Combinations
of a compound provided herein and other active ingredients will generally also be
within the aforementioned range, but in each case, an effective dose of each active
ingredient should be used.
4.5 CYCLING THERAPY
[0157] In certain embodiments, the prophylactic or therapeutic agents provided herein are
cyclically administered to a patient. Cycling therapy involves the administration
of an active agent for a period of time, followed by a rest for a period of time,
and repeating this sequential administration. Cycling therapy can reduce the development
of resistance to one or more of the therapies, avoid or reduce the side effects of
one of the therapies, and/or improves the efficacy of the treatment.
[0158] In certain embodiments, a compound provided herein is administered daily in a single
or divided dose in a four to six week cycle with a rest period of about a week or
two weeks. The disclosure further allows the frequency, number, and length of dosing
cycles to be increased. In certain embodiments, a compound provided herein is administered
for more cycles than are typical when it is administered alone. In certain embodiments,
a compound provided herein is administered for a greater number of cycles that would
typically cause dose-limiting toxicity in a patient to whom a second active ingredient
is not also being administered.
[0159] In one embodiment, a compound provided herein is administered daily and continuously
for three or four weeks at a dose of from about 0.1 mg to about 500 mg per day, followed
by a break of one or two weeks. In other embodiments, the dose can be from about 1
mg to about 300 mg, from about 0.1 mg to about 150 mg, from about 1 mg to about 200
mg, from about 10 mg to about 100 mg, from about 0.1 mg to about 50 mg, from about
1 mg to about 50 mg, from about 10 mg to about 50 mg, from about 20 mg to about 30
mg, or from about 1 mg to about 20 mg, followed by a break.
[0160] In one embodiment, a compound provided herein and a second active ingredient are
administered orally, with administration of the compound provided herein occurring
30 to 60 minutes prior to the second active ingredient, during a cycle of four to
six weeks. In another embodiment, the combination of a compound provided herein and
a second active ingredient is administered by intravenous infusion over about 90 minutes
every cycle. In certain embodiments, the number of cycles during which the combinatorial
treatment is administered to a patient is from about one to about 24 cycles, from
about two to about 16 cycles, or from about four to about three cycles.
4.6 PHARMACEUTICAL COMPOSITIONS AND DOSAGE FORMS
[0161] Pharmaceutical compositions can be used in the preparation of individual, single
unit dosage forms. Pharmaceutical compositions and dosage forms provided herein comprise
a compound provided herein, or a pharmaceutically acceptable salt, solvate, or stereoisomer
thereof. Pharmaceutical compositions and dosage forms provided herein can further
comprise one or more excipients.
[0162] Pharmaceutical compositions and dosage forms provided herein can also comprise one
or more additional active ingredients. Examples of optional second, or additional,
active ingredients are disclosed in Section 4.4, above.
[0163] Single unit dosage forms provided herein are suitable for oral, mucosal (
e.g., nasal, sublingual, vaginal, buccal, or rectal), parenteral (
e.g., subcutaneous, intravenous, bolus injection, intramuscular, or intraarterial), topical
(
e.g., eye drops or other ophthalmic preparations), transdermal or transcutaneous administration
to a patient. Examples of dosage forms include, but are not limited to: tablets; caplets;
capsules, such as soft elastic gelatin capsules; cachets; troches; lozenges; dispersions;
suppositories; powders; aerosols (
e.g., nasal sprays or inhalers); gels; liquid dosage forms suitable for oral or mucosal
administration to a patient, including suspensions (
e.g., aqueous or non-aqueous liquid suspensions, oil-in-water emulsions, or a water-in-oil
liquid emulsions), solutions, and elixirs; liquid dosage forms suitable for parenteral
administration to a patient; eye drops or other ophthalmic preparations suitable for
topical administration; and sterile solids (
e.g., crystalline or amorphous solids) that can be reconstituted to provide liquid dosage
forms suitable for parenteral administration to a patient.
[0164] The composition, shape, and type of dosage forms provided herein will typically vary
depending on their use. For example, a dosage form used in the acute treatment of
a disease may contain larger amounts of one or more of the active ingredients it comprises
than a dosage form used in the chronic treatment of the same disease. Similarly, a
parenteral dosage form may contain smaller amounts of one or more of the active ingredients
it comprises than an oral dosage form used to treat the same disease. These and other
ways in which specific dosage forms encompassed by this disclosure will vary from
one another will be readily apparent to those skilled in the art.
See, e.g., Remington's Pharmaceutical Sciences, 18th ed., Mack Publishing, Easton PA (1990).
[0165] In certain embodiments, the pharmaceutical compositions and dosage forms provided
herein comprise one or more excipients. Suitable excipients are well known to those
skilled in the art of pharmacy, and non-limiting examples of suitable excipients are
provided herein. Whether a particular excipient is suitable for incorporation into
a pharmaceutical composition or dosage form depends on a variety of factors well known
in the art including, but not limited to, the way in which the dosage form will be
administered to a patient. For example, oral dosage forms such as tablets may contain
excipients not suited for use in parenteral dosage forms. The suitability of a particular
excipient may also depend on the specific active ingredients in the dosage form. For
example, the decomposition of some active ingredients may be accelerated by some excipients
such as lactose, or when exposed to water. Active ingredients that comprise primary
or secondary amines are particularly susceptible to such accelerated decomposition.
Consequently, this disclosure encompasses pharmaceutical compositions and dosage forms
that contain little, if any, lactose other mono- or di-saccharides. As used herein,
the term "lactose-free" means that the amount of lactose present, if any, is insufficient
to substantially increase the degradation rate of an active ingredient.
[0166] Lactose-free compositions provided herein can comprise excipients that are well known
in the art and are listed, for example, in the
U.S. Pharmacopeia (USP) 25-NF20 (2002). In general, lactose-free compositions comprise active ingredients, a binder/filler,
and a lubricant in pharmaceutically compatible and pharmaceutically acceptable amounts.
In certain embodiments, lactose-free dosage forms provided herein comprise active
ingredients, microcrystalline cellulose, pre-gelatinized starch, and magnesium stearate.
[0167] This disclosure further encompasses anhydrous pharmaceutical compositions and dosage
forms comprising active ingredients, since water can facilitate the degradation of
some compounds. For example, the addition of water (
e.g., 5%) is widely accepted in the pharmaceutical arts as a means of simulating long-term
storage in order to determine characteristics such as shelf-life or the stability
of formulations over time.
See, e.g., Jens T. Carstensen, Drug Stability: Principles & Practice, 2d. Ed., Marcel Dekker,
NY, NY, 1995, pp. 379-80. In effect, water and heat accelerate the decomposition of some compounds. Thus,
the effect of water on a formulation can be of great significance since moisture and/or
humidity are commonly encountered during manufacture, handling, packaging, storage,
shipment, and use of formulations.
[0168] Anhydrous pharmaceutical compositions and dosage forms provided herein can be prepared
using anhydrous or low moisture containing ingredients and low moisture or low humidity
conditions. In certain embodiments, pharmaceutical compositions and dosage forms that
comprise lactose and at least one active ingredient that comprises a primary or secondary
amine are anhydrous if substantial contact with moisture and/or humidity during manufacturing,
packaging, and/or storage is expected.
[0169] An anhydrous pharmaceutical composition should be prepared and stored such that its
anhydrous nature is maintained. In certain embodiments, anhydrous compositions are
packaged using materials known to prevent exposure to water such that they can be
included in suitable formulary kits. Examples of suitable packaging include, but are
not limited to, hermetically sealed foils, plastics, unit dose containers (
e.g., vials), blister packs, and strip packs.
[0170] The disclosure further encompasses pharmaceutical compositions and dosage forms that
comprise one or more compounds that reduce the rate by which an active ingredient
will decompose. Such compounds, which are referred to herein as "stabilizers," include,
but are not limited to, antioxidants such as ascorbic acid, pH buffers, or salt buffers.
Like the amounts and types of excipients, the amounts and specific types of active
ingredients in a dosage form may differ depending on factors such as, but not limited
to, the route by which it is to be administered to patients. In certain embodiments,
dosage forms provided herein comprise a compound provided herein in an amount of from
about 0.10 to about 500 mg. In certain embodiments, the dosage forms provided herein
comprise a compound provided herein in an amount of about 0.1, 1, 2, 5, 7.5, 10, 12.5,
15, 17.5, 20, 25, 50, 100, 150, 200, 250, 300, 350, 400, 450, or 500 mg.
[0171] In certain embodiments, the dosage forms comprise the second active ingredient in
an amount of 1 to about 1,000 mg, from about 5 to about 500 mg, from about 10 to about
350 mg, or from about 50 to about 200 mg. Of course, the specific amount of the second
active agent will depend on the specific agent used, the type of diseases being treated
or managed, and the amount(s) of a compound provided herein and any optional additional
active agents concurrently administered to the patient.
4.6.1 ORAL DOSAGE FORMS
[0172] Pharmaceutical compositions provided herein that are suitable for oral administration
can be presented as discrete dosage forms, such as, but are not limited to, tablets
(
e.g., chewable tablets), caplets, capsules, and liquids (
e.g., flavored syrups). Such dosage forms contain predetermined amounts of active ingredients,
and may be prepared by methods of pharmacy well known to those skilled in the art.
See generally, Remington's Pharmaceutical Sciences, 18th ed., Mack Publishing, Easton PA (1990).
[0173] In certain embodiments, oral dosage forms provided herein are prepared by combining
the active ingredients in an intimate admixture with at least one excipient according
to conventional pharmaceutical compounding techniques. Excipients can take a wide
variety of forms depending on the form of preparation desired for administration.
For example, excipients suitable for use in oral liquid or aerosol dosage forms include,
but are not limited to, water, glycols, oils, alcohols, flavoring agents, preservatives,
and coloring agents. Examples of excipients suitable for use in solid oral dosage
forms (
e.g., powders, tablets, capsules, and caplets) include, but are not limited to, starches,
sugars, micro-crystalline cellulose, diluents, granulating agents, lubricants, binders,
and disintegrating agents. Because of their ease of administration, tablets and capsules
represent the most advantageous oral dosage unit forms, in which case solid excipients
are employed. If desired, tablets can be coated by standard aqueous or nonaqueous
techniques. Such dosage forms can be prepared by any of the methods of pharmacy. In
general, pharmaceutical compositions and dosage forms are prepared by uniformly and
intimately admixing the active ingredients with liquid carriers, finely divided solid
carriers, or both, and then shaping the product into the desired presentation if necessary.
[0174] For example, a tablet can be prepared by compression or molding. Compressed tablets
can be prepared by compressing in a suitable machine the active ingredients in a free-flowing
form such as powder or granules, optionally mixed with an excipient. Molded tablets
can be made by molding in a suitable machine a mixture of the powdered compound moistened
with an inert liquid diluent.
[0175] Examples of excipients that can be used in oral dosage forms provided herein include,
but are not limited to, binders, fillers, disintegrants, and lubricants. Binders suitable
for use in pharmaceutical compositions and dosage forms include, but are not limited
to, corn starch, potato starch, or other starches, gelatin, natural and synthetic
gums such as acacia, sodium alginate, alginic acid, other alginates, powdered tragacanth,
guar gum, cellulose and its derivatives (
e.g., ethyl cellulose, cellulose acetate, carboxymethyl cellulose calcium, sodium carboxymethyl
cellulose), polyvinyl pyrrolidone, methyl cellulose, pre-gelatinized starch, hydroxypropyl
methyl cellulose, (
e.g., Nos. 2208, 2906, 2910), microcrystalline cellulose, and mixtures thereof.
[0176] Suitable forms of microcrystalline cellulose include, but are not limited to, the
materials sold as AVICEL-PH-101, AVICEL-PH-103 AVICEL RC-581, AVICEL-PH-105 (available
from FMC Corporation, American Viscose Division, Avicel Sales, Marcus Hook, PA), and
mixtures thereof. An specific binder is a mixture of microcrystalline cellulose and
sodium carboxymethyl cellulose sold as AVICEL RC-581. Suitable anhydrous or low moisture
excipients or additives include AVICEL-PH-103™ and Starch 1500 LM.
[0177] Examples of fillers suitable for use in the pharmaceutical compositions and dosage
forms provided herein include, but are not limited to, talc, calcium carbonate (
e.g., granules or powder), microcrystalline cellulose, powdered cellulose, dextrates,
kaolin, mannitol, silicic acid, sorbitol, starch, pre-gelatinized starch, and mixtures
thereof. In certain embodiments, the binder or filler in pharmaceutical compositions
provided herein is present in from about 50 to about 99 weight percent of the pharmaceutical
composition or dosage form.
[0178] Disintegrants are used in the compositions provided herein to provide tablets that
disintegrate when exposed to an aqueous environment. Tablets that contain too much
disintegrant may disintegrate in storage, while those that contain too little may
not disintegrate at a desired rate or under the desired conditions. Thus, a sufficient
amount of disintegrant that is neither too much nor too little to detrimentally alter
the release of the active ingredients should be used to form solid oral dosage forms
provided herein. The amount of disintegrant used varies based upon the type of formulation,
and is readily discernible to those of ordinary skill in the art. In certain embodiments,
pharmaceutical compositions provided herein comprise from about 0.5 to about 15 weight
percent of disintegrant, or from about 1 to about 5 weight percent of disintegrant.
[0179] Disintegrants that can be used in pharmaceutical compositions and dosage forms provided
herein include, but are not limited to, agar-agar, alginic acid, calcium carbonate,
microcrystalline cellulose, croscarmellose sodium, crospovidone, polacrilin potassium,
sodium starch glycolate, potato or tapioca starch, other starches, pre-gelatinized
starch, other starches, clays, other algins, other celluloses, gums, and mixtures
thereof.
[0180] Lubricants that can be used in pharmaceutical compositions and dosage forms provided
herein include, but are not limited to, calcium stearate, magnesium stearate, mineral
oil, light mineral oil, glycerin, sorbitol, mannitol, polyethylene glycol, other glycols,
stearic acid, sodium lauryl sulfate, talc, hydrogenated vegetable oil (
e.g., peanut oil, cottonseed oil, sunflower oil, sesame oil, olive oil, corn oil, and soybean
oil), zinc stearate, ethyl oleate, ethyl laureate, agar, and mixtures thereof. Additional
lubricants include, for example, a syloid silica gel (AEROSIL200, manufactured by
W.R. Grace Co. of Baltimore, MD), a coagulated aerosol of synthetic silica (marketed
by Degussa Co. of Plano, TX), CAB-O-SIL (a pyrogenic silicon dioxide product sold
by Cabot Co. of Boston, MA), and mixtures thereof. If used at all, lubricants are
typically used in an amount of less than about 1 weight percent of the pharmaceutical
compositions or dosage forms into which they are incorporated.
[0181] A solid oral dosage form provided herein comprises a compound provided herein, anhydrous
lactose, microcrystalline cellulose, polyvinylpyrrolidone, stearic acid, colloidal
anhydrous silica, and gelatin.
4.6.2 CONTROLLED RELEASE DOSAGE FORMS
[0182] Active ingredients provided herein can be administered by controlled release means
or by delivery devices that are well known to those of ordinary skill in the art.
Examples include, but are not limited to, those described in
U.S. Patent Nos.: 3,845,770;
3,916,899;
3,536,809;
3,598,123; and
4,008,719,
5,674,533,
5,059,595,
5,591,767,
5,120,548,
5,073,543,
5,639,476,
5,354,556, and
5,733,566. Such dosage forms can be used to provide slow or controlled-release of one or more
active ingredients using, for example, hydropropylmethyl cellulose, other polymer
matrices, gels, permeable membranes, osmotic systems, multilayer coatings, microparticles,
liposomes, microspheres, or a combination thereof to provide the desired release profile
in varying proportions. Suitable controlled-release formulations known to those of
ordinary skill in the art, including those described herein, can be readily selected
for use with the active ingredients provided herein. The disclosure thus encompasses
single unit dosage forms suitable for oral administration such as, but not limited
to, tablets, capsules, gelcaps, and caplets that are adapted for controlled-release.
[0183] All controlled-release pharmaceutical products have a common goal of improving drug
therapy over that achieved by their non-controlled counterparts. Ideally, the use
of an optimally designed controlled-release preparation in medical treatment is characterized
by a minimum of drug substance being employed to cure or control the condition in
a minimum amount of time. Advantages of controlled-release formulations include extended
activity of the drug, reduced dosage frequency, and increased patient compliance.
In addition, controlled-release formulations can be used to affect the time of onset
of action or other characteristics, such as blood levels of the drug, and can thus
affect the occurrence of side (
e.g., adverse) effects.
[0184] Most controlled-release formulations are designed to initially release an amount
of drug (active ingredient) that promptly produces the desired therapeutic effect,
and gradually and continually release of other amounts of drug to maintain this level
of therapeutic or prophylactic effect over an extended period of time. In order to
maintain this constant level of drug in the body, the drug must be released from the
dosage form at a rate that will replace the amount of drug being metabolized and excreted
from the body. Controlled-release of an active ingredient can be stimulated by various
conditions including, but not limited to, pH, temperature, enzymes, water, or other
physiological conditions or compounds.
4.6.3 PARENTERAL DOSAGE FORMS
[0185] Parenteral dosage forms can be administered to patients by various routes including,
but not limited to, subcutaneous, intravenous (including bolus injection), intramuscular,
and intraarterial. Because their administration typically bypasses patients' natural
defenses against contaminants, parenteral dosage forms are preferably sterile or capable
of being sterilized prior to administration to a patient. Examples of parenteral dosage
forms include, but are not limited to, solutions ready for injection, dry products
ready to be dissolved or suspended in a pharmaceutically acceptable vehicle for injection,
suspensions ready for injection, and emulsions.
[0186] Suitable vehicles that can be used to provide parenteral dosage forms provided herein
are well known to those skilled in the art. Examples include, but are not limited
to: Water for Injection USP; aqueous vehicles such as, but not limited to, Sodium
Chloride Injection, Ringer's Injection, Dextrose Injection, Dextrose and Sodium Chloride
Injection, and Lactated Ringer's Injection; water-miscible vehicles such as, but not
limited to, ethyl alcohol, polyethylene glycol, and polypropylene glycol; and non-aqueous
vehicles such as, but not limited to, corn oil, cottonseed oil, peanut oil, sesame
oil, ethyl oleate, isopropyl myristate, and benzyl benzoate.
[0187] Compounds that increase the solubility of one or more of the active ingredients disclosed
herein can also be incorporated into the parenteral dosage forms provided herein.
For example, cyclodextrin and its derivatives can be used to increase the solubility
of an immunomodulatory compound provided herein and its derivatives.
See, e.g., U.S. Patent No. 5,134,127.
4.6.4 TOPICAL AND MUCOSAL DOSAGE FORMS
[0188] Topical and mucosal dosage forms provided herein include, but are not limited to,
sprays, aerosols, solutions, emulsions, suspensions, eye drops or other ophthalmic
preparations, or other forms known to one of skill in the art.
See, e.g., Remington's Pharmaceutical Sciences, 16th and 18th eds., Mack Publishing, Easton PA
(1980 & 1990); and
Introduction to Pharmaceutical Dosage Forms, 4th ed., Lea & Febiger, Philadelphia
(1985). Dosage forms suitable for treating mucosal tissues within the oral cavity can be
formulated as mouthwashes or as oral gels.
[0189] Suitable excipients (
e.g., carriers and diluents) and other materials that can be used to provide topical and
mucosal dosage forms encompassed by this disclosure are well known to those skilled
in the pharmaceutical arts, and depend on the particular tissue to which a given pharmaceutical
composition or dosage form will be applied. With that fact in mind, typical excipients
include, but are not limited to, water, acetone, ethanol, ethylene glycol, propylene
glycol, butane-1,3-diol, isopropyl myristate, isopropyl palmitate, mineral oil, and
mixtures thereof to form solutions, emulsions or gels, which are non-toxic and pharmaceutically
acceptable. Moisturizers or humectants can also be added to pharmaceutical compositions
and dosage forms if desired. Examples of such additional ingredients are well known
in the art.
See, e.g., Remington's Pharmaceutical Sciences, 16th and 18th eds., Mack Publishing, Easton PA
(1980 & 1990).
[0190] The pH of a pharmaceutical composition or dosage form may also be adjusted to improve
delivery of one or more active ingredients. Similarly, the polarity of a solvent carrier,
its ionic strength, or tonicity can be adjusted to improve delivery. Compounds such
as stearates can also be added to pharmaceutical compositions or dosage forms to advantageously
alter the hydrophilicity or lipophilicity of one or more active ingredients so as
to improve delivery. In this regard, stearates can serve as a lipid vehicle for the
formulation, as an emulsifying agent or surfactant, and as a delivery-enhancing or
penetration-enhancing agent. Different salts, hydrates or solvates of the active ingredients
can be used to further adjust the properties of the resulting composition.
4.7 KITS
[0191] The compounds provided herein can also be provided as an article of manufacture using
packaging materials well known to those of skill in the art.
See, e.g., U.S. Pat. Nos. 5,323,907;
5,052,558; and
5,033,252. Examples of pharmaceutical packaging materials include, but are not limited to,
blister packs, bottles, tubes, inhalers, pumps, bags, vials, containers, syringes,
and any packaging material suitable for a selected formulation and intended mode of
administration and treatment.
[0192] Provided herein also are kits which, when used by the medical practitioner, can simplify
the administration of appropriate amounts of active ingredients to a subject. In certain
embodiments, the kit provided herein includes a container and a dosage form of a compound
provided herein, including a single enantiomer, a mixture of an enantiomeric pair,
an individual diastereomer, or a mixture of diastereomers thereof; or a pharmaceutically
acceptable salt, solvate, or prodrug thereof.
[0193] In certain embodiments, the kit includes a container comprising a dosage form of
the compound provided herein, including a single enantiomer, a mixture of an enantiomeric
pair, an individual diastereomer, or a mixture of diastereomers thereof; or a pharmaceutically
acceptable salt, solvate, or prodrug thereof, in a container comprising one or more
other therapeutic agent(s) described herein. Kits provided herein can further comprise
additional active ingredients such as oblimersen (Genasense®), melphalan, G-CSF, GM-CSF,
EPO, topotecan, dacarbazine, irinotecan, taxotere, IFN, COX-2 inhibitor, pentoxifylline,
ciprofloxacin, dexamethasone, IL2, IL8, IL18, Ara-C, vinorelbine, isotretinoin, 13
cis-retinoic acid, or a pharmacologically active mutant or derivative thereof, or
a combination thereof. Examples of the additional active ingredients include, but
are not limited to, those disclosed herein.
[0194] Kits provided herein can further include devices that are used to administer the
active ingredients. Examples of such devices include, but are not limited to, syringes,
needle-less injectors drip bags, patches, and inhalers. The kits provided herein can
also include condoms for administration of the active ingredients.
[0195] Kits provided herein can further include pharmaceutically acceptable vehicles that
can be used to administer one or more active ingredients. For example, if an active
ingredient is provided in a solid form that must be reconstituted for parenteral administration,
the kit can comprise a sealed container of a suitable vehicle in which the active
ingredient can be dissolved to form a particulate-free sterile solution that is suitable
for parenteral administration. Examples of pharmaceutically acceptable vehicles include,
but are not limited to: aqueous vehicles, including, but not limited to, Water for
Injection USP, Sodium Chloride Injection, Ringer's Injection, Dextrose Injection,
Dextrose and Sodium Chloride Injection, and Lactated Ringer's Injection; water-miscible
vehicles, including, but not limited to, ethyl alcohol, polyethylene glycol, and polypropylene
glycol; and non-aqueous vehicles, including, but not limited to, corn oil, cottonseed
oil, peanut oil, sesame oil, ethyl oleate, isopropyl myristate, and benzyl benzoate.
[0196] The disclosure will be further understood by the following non-limiting examples.
5. EXAMPLES
5.1 1-[2-(2,6-Dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-3-(4-pyridin-4-ylmethyl-phenyl)-urea (Example disclosed for reference only)
[0197]

[0198] To a suspension of 3-(5-aminomethyl-1-oxo-1,3-dihydro-isoindol-2-yl)-piperidine-2,6-dione
methanesulfonic acid salt (0.18 g, 0.5 mmol) in DMF (2 mL) was added CDI (81 mg, 0.5
mmol). The mixture was stirred at RT overnight. 4-Pyridin-4-ylmethyl-phenylamine (92
mg, 0.5 mmol) was added to the mixture and the mixture was stirred at RT for 4 hrs.
Then the temperature was elevated to 40 °C and the mixture was stirred at this temperature
for 8 hrs. The mixture was cooled to RT, added water (5 mL), and stirred for 10 min.
The suspension was filtered and the solid was washed with water (20 mL), EtOAc (20
mL), and CH
3CN (20 mL) to give the product as an off-white solid (100 mg, 41% yield): HPLC: Waters
Symmetry C
18, 5 µm, 3.9 x 150 mm, 1 mL/min, 240 nm, 5% gradient 100% in 5 min, CH
3CN/0.1% H
3PO
4, 4.69 min (94%); mp: 290-292 °C;
1H NMR (DMSO-
d6)
δ 1.93 - 2.06 (m, 1H, CHH), 2.29 - 2.44 (m, 1H, CHH), 2.54 - 2.68 (m, 1H, CHH), 2.82
- 3.02 (m, 1H, CHH), 3.88 (s, 2H, CH
2), 4.24 - 4.54 (m, 4H, CH
2, CH
2), 5.10 (dd,
J = 5.0, 13.7 Hz, 1H, NCH), 6.70 (s, 1H, NH), 7.10 (d,
J = 8.3 Hz, 2H, Ar), 7.22 (d,
J = 5.3 Hz, 2H, Ar), 7.34 (d,
J = 8.3 Hz, 2H, Ar), 7.39 - 7.48 (m, 1H, Ar), 7.51 (s, 1H, Ar), 7.69 (d,
J = 7.7 Hz, 1H, Ar), 8.40 - 8.52 (m, 2H, Ar), 8.59 (s, 1H, NH), 10.98 (s, 1H, NH);
13C NMR (DMSO-
d6)
δ 22.49, 31.20, 40.63, 42.76, 47.12, 51.56, 118.03, 121.83, 122.91, 123.99, 126.86,
129.06, 130.27, 132.08, 138.75, 142.36, 144.87, 149.42, 155.22, 163.87, 167.93, 170.98,
172.85; LCMS MH
+ = 484; Anal. Calcd. for C
27H
25N
5O
4 + 1.5 H
2O: C, 63.52; H, 5.53; N, 13.72; S, 6.55; Found: C, 63.68; H, 5.24; N, 13.79.
5.2 1-[2-(2,6-Dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-3-(4-hydroxymethyl-phenyl)-urea (Example disclosed for reference only)
[0199]

[0200] To a suspension of 3-(5-aminomethyl-1-oxo-1,3-dihydro-isoindol-2-yl)-piperidine-2,6-dione
methanesulfonic acid salt (0.18 g, 0.5 mmol) in DMF (2 mL) was added CDI (81 mg, 0.5
mmol). The mixture was stirred at RT overnight. 4-Amino-benzyl alcohol (62 mg, 0.5
mmol) was added to the mixture and the mixture was stirred at RT for 4 hrs. Then the
temperature was elevated to 40 °C and the mixture was stirred at this temperature
for 8 hrs. The mixture was cooled to RT, added water (5 mL) and stirred for 10 min.
The suspension was filtered and the solid was washed with water (20 mL), EtOAc (20
mL), and CH
3CN (20 mL) to give a reddish solid (100 mg, 41% yield): The solid was purified on
ISCO silica gel column using methanol and DCM as eluent to give the product as a white
solid (30 mg, 15% yield): HPLC: Waters Symmetry C
18, 5 µm, 3.9 x 150 mm, 1 mL/min, 240 nm, 5% gradient 100% in 5 min, CH
3CN/0.1% H
3PO
4, 4.99 min (97%); mp: 309-311 °C;
1H NMR (DMSO-
d6)
δ 1.92 - 2.10 (m, 1H, CHH), 2.20 - 2.47 (m, 1H, CHH), 2.54 - 2.69 (m, 1H, CHH), 2.80
- 3.07 (m, 1H, CHH), 4.19 - 4.54 (m, 6H, CH
2, CH
2, CH
2), 4.94 - 5.05 (m, 1H, OH), 5.11 (dd,
J = 5.1, 13.2 Hz, 1H, CHN), 6.70 (t,
J = 6.0 Hz, 1H, NH), 7.16 (d,
J = 8.7 Hz, 2H, Ar), 7.28 - 7.40 (m, 2H, Ar), 7.39 - 7.48 (m, 1H, Ar), 7.52 (s, 1H,
Ar), 7.69 (d,
J = 7.7 Hz, 1H, Ar), 8.57 (s, 1H, NH), 10.98 (s, 1H, NH);
13C NMR (DMSO-
d6)
δ 22.49, 31.18, 42.77, 47.12, 51.56, 62.67, 117.49, 121.89, 122.93, 126.91, 127.05,
130.29, 135.22, 138.99, 142.38, 144.88, 155.25, 167.95, 170.99, 172.85; LCMS MH
+ = 423; Anal. Calcd. for C
22H
22N
4O
5 + 0.4 H
2O: C, 61.50; H, 5.35; N, 13.04; Found: C, 61.21; H, 5.05; N, 12.80.
5.3 1-[2-(2,6-Dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-3-[3-(2-methyl-2H-pyrazol-3-yl)-phenyl]-urea (Example disclosed for reference only)
[0201]

[0202] To a mixture of 5-(3-isocyanato-phenyl)-1-methyl-1
H-pyrazole (0.22 g, 1.1 mmol) and 3-(5-aminomethyl-1-oxo-1,3-dihydro-isoindol-2-yl)-piperidine-2,6-dione
methanesulfonic acid salt (0.40 g, 1.1 mmol) in acetonitrile (5 mL) was added TEA
(0.31 mL, 2.2 mmol) at RT, and the mixture was kept for 22 hrs. Water (25 mL) was
added to the mixture and the mixture was stirred at RT for 3 hrs. The suspension was
filtered and the solid was washed with water (20 mL), EtOAc (20 mL), and water (20
mL) to give a solid. The solid was purified with preparative HPLC to give the product
as a white solid (122 mg, 24% yield): HPLC: Waters Symmetry C
18, 5 µm, 3.9 x 150 mm, 1 mL/min, 240 nm, 30/70 CH
3CN/0.1% H
3PO
4, 3.34 min (99.8 %); mp: 260-262 °C;
1H NMR (DMSO-
d6)
δ 1.88 - 2.10 (m, 1H, CHH), 2.27 - 2.47 (m, 1H, CHH), 2.55 - 2.66 (m, 1H, CHH), 2.80
- 3.02 (m, 1H, CHH), 3.84 (s, 3H, CH
3), 4.30 (d,
J = 17.6 Hz, 1H, CHH), 4.40 - 4.50 (m, 3H, CHH, CH
2), 5.11 (dd,
J = 5.2, 13.3 Hz, 1H, NCH), 6.35 (d,
J = 1.9 Hz, 1H, Ar), 6.83 (t,
J = 6.1 Hz, 1H, NH), 7.06 (dt,
J = 1.4, 7.6 Hz, 1H, Ar), 7.31 - 7.38 (m, 1H, Ar), 7.40 - 7.48 (m, 3H, Ar), 7.53 (d,
J = 0.4 Hz, 1H, Ar), 7.63 (t,
J = 1.9 Hz, 1H, Ar), 7.70 (d,
J = 7.9 Hz, 1H, Ar), 8.82 (s, 1H, NH), 10.98 (br. s., 1H, NH);
13C NMR (DMSO-
d6) δ 22.48, 31.19, 37.45, 42.78, 47.11, 51.55, 105.55, 117.62 (2 carbons by HMQC),
121.18, 121.87, 122.93, 126.87, 129.08, 130.29, 130.51, 137.86, 140.73, 142.38, 142.82,
144.76, 155.21, 167.94, 170.98, 172.84; LCMS MH
+ = 473; Anal. Calcd. for C
25H
24N
6O
4: C, 63.55; H, 5.12; N, 17.79; Found: C, 63.36; H, 5.17; N, 17.72.
5.4 1-[2-(2,6-Dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-3-[3-(2-methyl-imidazol-1-yl)-phenyl]-urea; formic acid (Example disclosed for reference only)
[0203]

[0204] A mixture of 3-(5-aminomethyl-1-oxo-1,3-dihydro-isoindol-2-yl)-piperidine-2,6-dione
methanesulfonic acid salt (0.40 g, 1.1 mmol) and CDI (0.19 g, 1.2 mmol) in DMF (5
mL) was stirred at RT for 18 hrs. To the mixture was added 3-(2-methyl-imidazol-1-yl)-phenylamine
(0.19 g, 1.1 mmol) at RT, and the mixture was stirred at 60 °C for 24 hrs. To the
mixture was added water (25 mL) and ether (20 mL). The mixture was stirred at RT for
2 hrs. The suspension was filtered and the solid was washed with water (20 mL), ethyl
acetate (20 mL), and water (20 mL) to give a solid. The solid was purified with preparative
HPLC to give the product as a white solid (100 mg, 20% yield): HPLC: Waters Symmetry
C
18, 5 µm, 3.9 x 150 mm, 1 mL/min, 240 nm, 5% gradient 95% in 5 min, CH
3CN/0.1% H
3PO
4, 4.48 min (96.8 %); mp: 218-220 °C;
1H NMR (DMSO-
d6)
δ 1.89 - 2.12 (m, 1H, CHH), 2.25 - 2.29 (m, 3H, CH
3), 2.30 - 2.46 (m, 1H, CHH), 2.54 - 2.68 (m, 1H, CHH), 2.78 - 3.04 (m, 1H, CHH), 4.31
(d,
J = 17.4 Hz, 1H, CHH), 4.38 - 4.53 (m, 3H, CHH, CH
2), 5.11 (dd,
J = 5.1, 13.2 Hz, 1H, NCH), 6.89 (d,
J = 1.3 Hz, 1H, Ar), 6.92 - 7.05 (m, 2H, Ar, NH), 7.23 (d,
J = 1.3 Hz, 1H, Ar), 7.32 - 7.40 (m, 2H, Ar), 7.41 - 7.49 (m, 1H, Ar), 7.52 (s, 1H,
Ar), 7.57 - 7.65 (m, 1H, Ar), 7.69 (d,
J = 7.9 Hz, 1H, Ar), 8.18 (s, 1H, HCOOH), 9.03 (s, 1H, NH), 10.98 (s, 1H, NH);
13C NMR (DMSO-
d6)
δ 13.60, 22.49, 31.20, 42.77, 47.12, 51.58, 114.22, 116.89, 117.61, 120.68, 121.88,
122.93, 126.89, 127.13, 129.63, 130.30, 137.90, 141.50, 142.38, 143.43, 144.69, 155.16,
163.44, 167.93, 170.98, 172.85; LCMS MH
+ = 473; Anal. Calcd. for C
25H
24N
6O
4 + HCOOH + 1.5 H
2O: C, 57.24; H, 5.36; N, 15.40; Found: C, 57.43; H, 5.11; N, 15.57.
5.5 1-[2-(2,6-Dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-3-[4-(4-methyl-4H-[1,2,4]triazol-3-yl)-phenyl]-urea (Example disclosed for reference only)
[0205]

[0206] To a stirred suspension of 4-(4-methyl-4
H-[1,2,4]triazol-3-yl)-phenylamine (0.31 g, 1.77 mmol) in DMF (10 mL) at 40 °C was added
CDI (0.32 g, 1.94 mmol). The mixture was stirred for 15 min, followed by addition
of 3-(5-aminomethyl-1-oxo-1,3-dihydro-isoindol-2-yl)-piperidine-2,6-dione methanesulfonic
acid salt (0.65 g, 1.77 mmol). Heating was stopped after 1.5 hrs and the mixture was
stirred at RT overnight. Solvent was evaporated and the residue was purified by preparative
HPLC to give the product as a white solid (0.13 g, 15% yield): HPLC, Waters Symmetry
C
18, 5 µm, 3.9 x 150 mm, 1 ml/min, 240 nm, 14/86 CH
3CN/0.1% H
3PO
4, 4.70 min (93.9%); mp, 248-250 °C;
1H NMR (DMSO-
d6)
δ 1.93 - 2.06 (m, 1H, CH
H), 2.29 - 2.46 (m, 1H, CH
H), 2.55 - 2.70 (m, 1H, CH
H), 2.82 - 3.02 (m, 1H, CH
H), 3.72 (s, 3H, C
H3), 4.24 - 4.55 (m, 4H, C
H2, C
H2), 5.11 (dd,
J = 4.9, 13.2 Hz, 1H, N
CH), 6.95 (t,
J = 5.7 Hz, 1H, N
H), 7.41 - 7.81 (m, 7H, Ar
H), 8.51 (s, 1H, Ar
H), 9.02 (s, 1H, N
H), 10.99 (s, 1H, N
H);
13C NMR (DMSO-
d6)
δ 22.49, 31.20, 31.94, 42.79, 47.13, 51.58, 117.50, 119.49, 121.88, 122.94, 126.91,
128.80, 130.32, 141.84, 142.39, 144.72, 145.76, 153.08, 155.12, 167.95, 170.99, 172.85;
LC/MS MH
+ = 474; Anal. Calcd. For C
24H
23N
7O
4: C, 60.88; H, 4.90; N, 20.71. Found: C, 58.28; H, 4.67; N, 19.49 (Note: This analysis
was off, and both HPLC and
1H NMR showed 6% impurity).
5.6 1-[2-(2,6-Dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-3-[3-(2-methyl-thiazol-4-yl)-phenyl]-urea (Example disclosed for reference only)
[0207]

[0208] To a mixture of 4-(3-isocyanato-phenyl)-2-methyl-thiazole (0.25 g, 1.2 mmol) and
3-(5-aminomethyl-1-oxo-1,3-dihydro-isoindol-2-yl)-piperidine-2,6-dione methanesulfonic
acid salt (0.43 g, 1.2 mmol) in acetonitrile (5 mL) was added TEA (0.33 mL, 2.3 mmol)
at RT, and the mixture was kept for 3 hrs. Water (25 mL) was added to the mixture
and the mixture was stirred at RT for 3 hrs. The suspension was filtered and the solid
was washed with water (20 mL), ethyl acetate (20 mL), and water (20 mL) to give a
solid. The solid was purified with preparative HPLC to give the product as a white
solid (160 mg, 28% yield): HPLC: Waters Symmetry C
18, 5 µm, 3.9 x 150 mm, 1 mL/min, 240 nm, 30/70 CH
3CN/0.1% H
3PO
4, 5.93 min (99.4 %); mp: 252-254 °C;
1H NMR (DMSO-
d6)
δ 1.92 - 2.06 (m, 1H, CHH), 2.27 - 2.47 (m, 1H, CHH), 2.54 - 2.66 (m, 1H, CHH), 2.71
(s, 3H, C
H3), 2.81 - 3.01 (m, 1H, CHH), 4.31 (d,
J = 17.6 Hz, 1H, CHH), 4.38 - 4.53 (m, 3H, CHH, CH
2), 5.11 (dd,
J = 5.1, 13.2 Hz, 1H, NCH), 6.74 (t,
J = 6.0 Hz, 1H, NH), 7.20 - 7.32 (m, 1H, Ar), 7.40 (ddd,
J = 1.1, 2.3, 8.1 Hz, 1H, Ar), 7.46 (dq,
J = 1.4, 7.6 Hz, 2H, Ar), 7.53 (s, 1H, Ar), 7.70 (d,
J = 7.9 Hz, 1H, Ar), 7.81 (s, 1H, Ar), 8.02 (t,
J = 1.9 Hz, 1H, Ar), 8.79 (s, 1H, NH), 10.98 (s, 1H, NH);
13C NMR (DMSO-
d6)
δ 18.91, 22.51, 31.18, 42.79, 47.12, 51.56, 113.55, 115.55, 117.28, 118.86, 121.86,
122.94, 126.89, 129.01, 130.29, 134.58, 140.82, 142.39, 144.90, 153.89, 155.21, 165.29,
167.95, 170.99, 172.85; LCMS MH
- = 490; Anal. Calcd. for C
25H
23N
5O
4S: C, 61.34; H, 4.74; N, 14.31; S, 6.55; Found: C, 61.09; H, 4.60; N, 14.19; S, 6.49.
5.7 1-[2-(2,6-Dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-3-[3-(1-methyl-1H-pyrazol-3-yl)-phenyl]-urea (Example disclosed for reference only)
[0209]

[0210] To a stirred suspension of 3-(3-isocyanato-phenyl)-1-methyl-1
H-pyrazole (0.25 g, 1.25 mmol) and 3-(5-aminomethyl-1-oxo-1,3-dihydro-isoindol-2-yl)-piperidine-2,6-dione
methanesulfonic acid salt (0.46 g, 1.25 mmol) in acetonitrile (5 mL) at RT was added
TEA (0.35 mL, 2.51 mmol). The mixture was stirred for 4 hrs, followed by addition
of 1N HCl (10 mL), which was stirred for 10 min. The mixture was purified by preparative
HPLC to give the product as an off-white solid (0.22 g, 38% yield): HPLC, Waters Symmetry
C
18, 5 µm, 3.9 x 150 mm, 1 ml/min, 240 nm, 25/75 CH
3CN/0.1% H
3PO
4, 5.99 min (99.9%); mp, 232-234 °C;
1H NMR (DMSO-
d6)
δ 1.93 - 2.07 (m, 1H, CH
H), 2.29 - 2.46 (m, 1H, CH
H), 2.55 - 2.68 (m, 1H, CH
H), 2.82 - 3.01 (m, 1H, CH
H), 3.87 (s, 3H, C
H3), 4.24 - 4.55 (m, 4H, C
H2, C
H2), 5.11 (dd,
J = 5.0, 13.3 Hz, 1H, NC
H), 6.57 (d,
J = 2.3 Hz, 1H, Ar
H), 6.73 (t,
J = 5.9 Hz, 1H, N
H), 7.18 - 7.37 (m, 3H, Ar
H), 7.46 (d,
J = 7.9 Hz, 1H, Ar
H), 7.53 (s, 1H, Ar
H), 7.65 - 7.77 (m, 2H, Ar
H), 7.89 (s, 1H, Ar
H), 8.71 (s, 1H, N
H), 10.98 (s, 1H, N
H);
13C NMR (DMSO-
d6)
δ 22.51, 31.18, 38.60, 42.79, 47.13, 51.58, 102.33, 114.31, 116.77, 118.16, 121.86,
122.93, 126.89, 128.85, 130.29, 132.17, 133.85, 140.66, 142.39, 144.91, 150.03, 155.22,
167.95, 170.99, 172.85; LC/MS MH
+ = 473; Anal. Calcd. For C
25H
24N
6O
4 +0.5 H
2O: C, 62.36; H, 5.23; N, 17.45. Found: C, 62.06; H, 5.19; N, 17.28.
5.8 1-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-3-(3-(morpholinomethyl)phenyl)urea
formate (Example disclosed for reference only)
[0211]

[0212] To a stirred mixture of 3-(5-aminomethyl-1-oxo-1,3-dihydro-isoindol-2-yl)-piperidine-2,6-dione
hydrochloride (0.37 g, 1.00 mmol) and 4-(3-isocyanatobenzyl)-morpholine (0.22 g, 1.00
mmol) in acetonitrile (10 mL) was added TEA (0.28 mL, 2.00 mmol) at RT under nitrogen.
After 2 hrs, additional 4-(3-isocyanatobenzyl)morpholine (0.22 g, 1.00 mmol) and TEA
(0.28 mL, 2.00 mmol) were added. After 12 hrs, undesired solid was filtered and filtrate
was concentrated. The residue was dissolved in acetonitrile and was purified by preparative
HPLC (gradient: CH
3CN + 0.1% formic acid/H
2O + 0.1% formic acid: 10/90 for 5 min, to 100/0 in 10 min, 100/0 for 5 min). After
evaporation of the solvent, the residue was triturated in ether (20 mL) for 1 hr.
The product was then isolated by filtration and dried
in vacuo to give the product as a beige solid (0.16 g, 30% yield): HPLC: Waters Symmetry C
18, 5 µm, 3.9 x 150 mm, 1 mL/min, 240 nm, gradient: CH
3CN/0.1% H
3PO
4: 10/90 to 90/10 in 10 min, 90/10 (5 min): 4.64 min (95.84%); mp: 198-200 °C;
1H NMR (DMSO-
d6)
δ 1.83 - 2.12 (m, 1H, CHH), 2.21 - 2.47 (m, 5H, CHH, CH
2, CH
2), 2.54 - 2.69 (m,
J = 11.0 Hz, 1H, CHH), 2.79 - 3.03 (m, 1H, CHH), 3.38 (s, 2H, CH
2), 3.52 - 3.72 (m, 4H, CH
2, CH
2), 4.31 (d,
J = 17.2 Hz, 1H, CHH), 4.37 - 4.55 (m, 3H, CHH, CH
2), 5.11 (dd,
J = 5.1, 13.2 Hz, 1H, CH), 6.83 (d,
J = 7.6 Hz, 1H, Ar), 6.99 (t,
J = 5.6 Hz, 1H, NH), 7.15 (t,
J = 7.7 Hz, 1H, Ar), 7.32 (d,
J = 8.3 Hz, 1H, Ar), 7.39 (s, 1H, Ar), 7.44 (d,
J = 7.7 Hz, 1H, Ar), 7.52 (s, 1H, Ar), 7.69 (d,
J = 7.7 Hz, 1H, Ar), 8.29 (br. s., 1H, HCOO), 8.90 (s, 1H, NH), 10.98 (br. s., 1H,
NH);
13C NMR (DMSO-
d6)
δ 22.51, 31.20, 42.73, 47.12, 51.56, 53.20, 62.69, 66.17, 116.42, 118.13, 121.72, 121.83,
122.90, 126.86, 128.34, 130.25, 138.24, 140.48, 142.36, 144.99, 155.31, 164.30, 167.95,
170.98, 172.85; LCMS: MH
+ = 492; Anal. Calcd. for C
27H
31N
5O
7 + 3 H
2O: C, 54.82; H, 6.30; N, 11.84; Found: C, 55.12; H, 6.12; N, 11.72.
5.9 1-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-3-(4-methyl-3-nitrophenyl)urea (Example disclosed for reference only)
[0213]

[0214] To a stirred mixture of 3-(5-aminomethyl-1-oxo-1,3-dihydro-isoindol-2-yl)-piperidine-2,6-dione
hydrochloride (1.00 g, 2.70 mmol) and 4-methyl-3-nitrophenylisocyanate (0.48 mL, 2.70
mmol) in acetonitrile (20 mL) was added TEA (0.75 mL, 5.40 mmol) at RT under nitrogen.
After 2 hrs, 1N HCl (20 mL) was added and the solids were isolated by filtration and
washed with water (3 x 20 mL). The crude product was triturated in EtOAc (50 mL) for
12 hrs. The product was isolated by filtration and dried
in vacuo to give the product as a yellow solid (0.74 g, 61% yield): HPLC: Waters Symmetry
C
18, 5 µm, 3.9 x 150 mm, 1 mL/min, 240 nm, gradient: CH
3CN/0.1% H
3PO
4: 10/90 to 90/10 in 10 min, 90/10 (5 min): 7.94 min (96.79%); mp: 230-232 °C;
1H NMR (DMSO-
d6)
δ 1.87 - 2.08 (m, 1H, CHH), 2.26 - 2.48 (m, 3H, CHH, C
H3), 2.60 (d,
J = 17.6 Hz, 1H, CHH), 2.79 - 3.07 (m, 1H, CHH), 4.31 (d,
J = 17.4 Hz, 1H, CHH), 4.38 - 4.55 (m, 3H, CHH, CH
2), 5.11 (dd,
J = 5.0, 13.3 Hz, 1H, CH), 6.92 (t,
J = 5.9 Hz, 1H, NH), 7.34 (d,
J = 8.3 Hz, 1H, Ar), 7.45 (d,
J = 7.9 Hz, 1H, Ar), 7.49 - 7.60 (m, 2H, Ar), 7.70 (d,
J = 7.7 Hz, 1H, Ar), 8.26 (d,
J = 2.3 Hz, 1H, Ar), 9.07 (s, 1H, NH), 10.99 (s, 1H, NH);
13C NMR (DMSO-
d6)
δ 18.95, 22.49, 31.20, 42.82, 47.12, 51.56, 112.74, 121.92, 122.45, 122.94, 124.71,
126.92, 130.33, 132.84, 139.46, 142.38, 144.59, 148.72, 155.02, 167.93, 170.98, 172.85;
LCMS: MH
+= 452; Anal. Calcd. for C
22H
21N
4O
6: C, 58.53; H, 4.69; N, 15.51; Found: C, 58.23; H, 4.58; N, 15.34.
5.10 1-(3-amino-4-methylphenyl)-3-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)urea (Example disclosed for reference only)
[0215]

[0216] To a solution of 1-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-3-(4-methyl-3-nitrophenyl)urea
(0.30 g, 0.66 mmol) in DMF (50 mL) was added Pd-C (0.10 g, 10% weight). The reaction
mixture was hydrogenated with a Parr-shaker at 55 psi. After 16 hrs, the mixture was
filtered through a celite pad, which was washed with additional DMF (20 mL). The filtrate
was then evaporated and the residue was stirred in water (100 mL) for 3 hrs. The solid
was filtered, washed with additional water (50 mL), and dried. The crude green product
was dissolved in DMF (100 mL), decolorizing carbon was added, and the reaction mixture
was stirred for 3 hrs. The mixture was then filtered through a celite pad, which was
washed with additional DMF (50 mL). The filtrate was then evaporated and the residue
was stirred in water (100 mL) for 4 hrs. The solid was filtered, washed with additional
water (50 mL), and dried
in vacuo. The solid was triturated with ether for 1 hr and the product was isolated by filtration
to give the product as a pale green solid (0.22 g, 79% yield): HPLC: Waters Symmetry
C
18, 5 µm, 3.9 x 150 mm, 1 mL/min, 240 nm, gradient: CH
3CN/0.1% H
3PO
4: 10/90 to 90/10 in 10 min, 90/10 (5 min): 4.50 min (96.74%); mp: 228-230 °C;
1H NMR (DMSO-
d6)
δ 1.82 - 2.10 (m, 4H, CHH, CH
3), 2.25 - 2.47 (m, 1H, CHH), 2.59 (d,
J = 18.3 Hz, 1H, CHH), 2.77 - 3.02 (m, 1H, CHH), 4.30 (d,
J = 17.6 Hz, 1H, CHH), 4.35 - 4.53 (m, 3H, CH
2, CHH), 4.93 (br. s., 2H, NH
2), 5.11 (dd,
J = 5.0, 12.9 Hz, 1H, CH), 6.51 (dd,
J = 1.9, 7.9 Hz, 1H, Ar), 6.58 (t,
J = 5.7 Hz, 1H, NH), 6.75 (d,
J = 8.5 Hz, 2H, Ar), 7.43 (d,
J = 7.9 Hz, 1H, Ar), 7.50 (s, 1H, Ar), 7.69 (d,
J = 7.7 Hz, 1H, Ar), 8.23 (s, 1H, NH), 10.98 (s, 1H, NH);
13C NMR (DMSO-
d6)
δ 16.75, 22.49, 31.20, 42.74, 47.10, 51.56, 104.16, 106.57, 114.59, 121.86, 122.90,
126.89, 129.79, 130.25, 138.75, 142.36, 145.06, 146.12, 155.22, 167.95, 170.98, 172.85;
LCMS: MH
+ = 422; Anal. Calcd. for C
22H
23N
5O
4 + 0.1 H
2O + 0.5 Et
2O: C, 62.62; H, 6.17; N, 15.21; Found: C, 62.30; H, 5.89; N, 14.89.
5.11 1-[2-(2,6-Dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-3-(3-phenoxy-phenyl)-urea (Example disclosed for reference only)
[0217]

[0218] TEA (0.20 g, 2.00 mmol) was added to a stirred mixture of 3-(5-aminomethyl-1-oxo-1,3-dihydro-isoindol-2-yl)-piperidine-2,6-dione
methanesulfonate (0.37 g, 1.00 mmol) and 1-isocyanato-3-phenoxy-benzene (0.212 g,
1.00 mmol) in acetonitrile (10 mL) under nitrogen at RT. After 3 hrs, 1N HCl (10 mL)
was added, and the mixture was stirred for 10 min. The solids were isolated by filtration,
and washed with water (20 mL) and acetonitrile (10 mL). The crude product was dissolved
in a minimal amount of DMF and the product was precipitated by slow addition of water
(∼30 mL). The solids were collected by filtration, washed with Et
2O, and dried
in vacuo for 18 hrs to give the product as a white solid (422 mg, 87%): HPLC: Waters Symmetry
C
18, 5 µm, 3.9 x 150 mm, 1 ml/min, 240 nm, 35/65, CH
3CN/0.1% H
3PO
4, 3.76 min (99.3%); mp: 224-226 °C;
1H NMR (DMSO-
d6)
δ 1.99 (s, 4H, CH, CH
3), 2.29 - 2.46 (m, 1H, CH), 2.55 - 2.66 (m, 1H, CH), 2.80 - 3.01 (m, 1H, CH), 4.18
- 4.56 (m, 4H, CH
2, CH
2), 5.10 (dd,
J = 4.9, 13.2 Hz, 1H, CH), 6.71 (t,
J = 5.9 Hz, 1H, NH), 7.23 - 7.36 (m, 1H, Ar), 7.36 - 7.48 (m, 2H, Ar), 7.51 (s, 1H,
Ar), 7.69 (d,
J = 7.7 Hz, 1H, Ar), 8.55 (s, 1H, NH), 9.75 (s, 1H, NH), 10.67 - 11.27 (m, 1H, NH);
13C NMR (DMSO-
d6)
δ 22.49, 23.82, 31.18, 42.77, 47.12, 51.56, 118.12, 119.57, 121.85, 122.91, 126.88,
130.26, 133.18, 135.70, 142.36, 144.94, 155.29, 167.67, 167.95, 170.98, 172.85; LCMS:
MH
+ = 485; Anal. Calcd. for C
27H
24N
4O
5: C, 66.93; H, 4.99; N, 11.56. Found: C, 67.03; H, 4.72; N, 11.41.
5.12 1-((2-(2,6-Dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-3-(4-nitrophenyl)urea (Example disclosed for reference only)
[0219]

[0220] TEA (0.20 g, 2.0 mmol) was added to a stirred mixture of 3-(5-aminomethyl-1-oxo-1,3-dihydro-isoindol-2-yl)-piperidine-2,6-dione
methanesulfonate (0.37 g, 1.0 mmol) and 1-isocyanato-4-nitrobenzene (164 mg, 1.0 mmol)
in acetonitrile (10 mL) under nitrogen. The mixture stirred at RT for 3 hrs, during
which time it remained as a suspension. A 1N HCl solution (10 mL) was added, and the
mixture was stirred for 10 min. The solid was isolated by filtration and washed with
additional water (20 mL) and acetonitrile (10 mL). The solid was dissolved in minimal
amount of DMF and the product was precipitated by slow addition of water (∼30 mL).
The solid was collected by filtration and washed with Et
2O to remove most of the residual yellow color. The remaining solid was dried in a
vacuum oven overnight to provide the product as an off-white solid (330 mg, 75%):
HPLC: Waters Symmetry C
18, 5 µm, 3.9 x 150 mm, 1 ml/min, 240 nm, 30/70, CH
3CN/0.1% H
3PO
4, 6.15 min (97.1%); mp: 272-274 °C;
1H NMR (DMSO-
d6)
δ 1.84 - 2.13 (m, 1H, CHH), 2.24 - 2.44 (m, 1H, CHH), 2.55 - 2.66 (m, 1H, CHH), 2.77
- 3.02 (m, 1H, CHH), 4.16 - 4.55 (m, 4H, CH
2, CH
2), 5.11 (dd,
J = 4.7, 13.0 Hz, 1H, CH), 7.07 (t,
J = 5.6 Hz, 1H, NH), 7.37 - 7.83 (m, 5H, Ar), 8.15 (d,
J = 8.9 Hz, 2H, Ar), 9.48 (s, 1H, NH), 10.98 (s, 1H, NH);
13C NMR (DMSO-
d6)
δ 22.48, 31.17, 42.83, 47.11, 51.57, 116.97, 121.94, 122.96, 125.09, 126.93, 130.38,
140.48, 142.41, 144.26, 147.04, 154.51, 167.91, 170.98, 172.85; LCMS: MH
+ = 438; Anal. Calcd. for C
21H
19N
5O
6 + 0.5 H
2O: C, 56.50; H, 4.52; N, 15.69; Found: C, 56.45; H, 4.31; N, 15.71.
5.13 N-(4-{3-[2-(2,6-Dioxo-piperidin-3-yl)-1-oxo-2,3-dihvdro-1H-isoindol-5-ylmethyl]-ureido}-pheny)-acetamide (Example disclosed for reference only)
[0221]

[0222] Step 1: Preparation of 1-(4-amino-phenyl)-3-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1
H-isoindol-5-ylmethyl]-urea. To a stirred mixture of 1-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-3-(4-nitrophenyl)urea
(150 mg, 0.343 mmol) in EtOH (2 mL) was added sodium dithionite (597 mg, 3.43 mmol)
in water (2 mL). The resulting mixture was heated to 60 °C for 20 min at which time
LC-MS indicated complete disappearance of nitro starting material. The reaction mixture
was combined with the crude product from a separate run and concentrated
in vacuo. The residue was dissolved in minimal DMF and chromatographed on a C-18 preparative
HPLC column equipped with mass triggered collection. The desired fractions were combined
and concentrated
in vacuo to provide 1-(4-amino-phenyl)-3-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1
H-isoindol-5-ylmethyl]-urea as a pale yellow solid (90 mg, 40% combined average yield
from two separate runs): mp: >400 °C; LCMS: MH
+ = 408.
[0223] Step 2: Preparation of
N-(4-{3-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1
H-isoindol-5-ylmethyl]-ureido}-phenyl)-acetamide. 1-(4-Amino-phenyl)-3-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1
H-isoindol-5-ylmethyl]-urea (64 mg, 0.157 mmol) was stirred at room temperature in
acetic anhydride (5 mL) for 2 hrs. The volatiles were removed
in vacuo and the residue was dissolved in minimal DMF and purified on a C-18 preparative HPLC
column. The desired fractions were combined and concentrated
in vacuo to provide the product as a pale yellow solid (37 mg, 52% yield): HPLC: Waters Symmetry
C
18, 5 µm, 3.9 x 150 mm, 1 ml/min, 240 nm, 17/83, CH
3CN/0.1% H
3PO
4, 5.19 min (97.4%); mp: 265-267 °C;
1H NMR (DMSO-
d6) δ 1.99 (s, 4H, CH,CH
3), 2.29 - 2.46 (m, 1H, CH), 2.55 - 2.66 (m, 1H, CH), 2.80 - 3.01 (m, 1H, CH), 4.18
- 4.56 (m, 4H, CH
2, CH
2), 5. 10 (dd,
J = 4.9, 13.2 Hz, 1H, CH), 6.71 (t,
J = 5.9 Hz, 1H, NH), 7.23 - 7.36 (m, 1H, Ar), 7.36 - 7.48 (m, 2H, Ar), 7.51 (s, 1H,
Ar), 7.69 (d,
J = 7.7 Hz, 1H, Ar), 8.55 (s, 1H, NH), 9.75 (s, 1H, NH), 10.67 - 11.27 (m, 1H, NH);
13C NMR (DMSO-
d6)
δ 22.49, 23.82, 31.18, 42.77, 47.12, 51.56, 118.12, 119.57, 121.85, 122.91, 126.88,
130.26, 133.18, 135.70, 142.36, 144.94, 155.29, 167.67, 167.95, 170.98, 172.85; LCMS:
MH
+ = 450; Anal. Calcd. for C
23H
23N
5O
5 + 1.0 H
2O: C, 59.09; H, 5.39; N, 14.98; Found: C, 58.75; H, 4.99; N, 14.59.
5.14 3-[2-(2,6-Dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-1-methyl-1-phenyl-urea (Example disclosed for reference only)
[0224]

[0225] To a suspension of 3-(5-aminomethyl-1-oxo-1,3-dihydro-isoindol-2-yl)-piperidine-2,6-dione
methanesulfonic acid salt (0.18 g, 0.5 mmol) in CH
3CN (10 mL) was added DIPEA (0.4 mL, 2.5 mmol) and
N-methylphenyl carbamic chloride (178 mg, 1.05 mmol). The mixture was stirred at RT
overnight. The suspension was filtered and the solid was washed with water (20 mL),
ethyl acetate (20 mL), and CH
3CN (20 mL) to give the product as a white solid (200 mg, 47% yield): HPLC: Waters
Symmetry C
18, 5 µm, 3.9 x 150 mm, 1 mL/min, 240 nm, 30/70 CH
3CN/0.1% H
3PO
4, 3.56 min (96 %); mp: 168-170 °C;
1H NMR (DMSO-
d6)
δ 1.89 - 2.07 (m, 1H, CHH), 2.30 - 2.47 (m, 1H, CHH), 2.55 - 2.69 (m, 1H, CHH), 2.79
- 3.01 (m, 1H, CHH), 3.18 (s, 3H, CH
3), 4.22 - 4.58 (m, 4H, CH
2, CH
2), 5.11 (dd,
J = 5.0, 13.3 Hz, 1H, NCH), 6.68 (t,
J = 5.9 Hz, 1H, NH), 7.16 - 7.35 (m, 3H, Ar), 7.36 - 7.44 (m, 3H, Ar), 7.46 (s, 1H,
Ar), 7.66 (d,
J = 7.7 Hz, 1H, Ar), 10.98 (s, 1H, NH);
13C NMR (DMSO-
d6)
δ 22.51, 31.21, 37.14, 43.76, 47.10, 51.55, 121.85, 122.71, 125.78, 126.54, 126.86,
129.23, 130.04, 142.19, 144.02, 145.41, 156.74, 168.02, 171.02, 172.86; LCMS MH
+= 407; Anal. Calcd. for C
22H
22N
4O
4+ 0.5 H
2O: C, 63.60; H, 5.58; N, 13.49; S, 6.55; Found: C, 63.61; H, 5.51; N, 13.48.
5.15 1-Biphenyl-4-yl-3-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-urea (Example disclosed for reference only)
[0226]

[0227] To a suspension of 3-(5-aminomethyl-1-oxo-1,3-dihydro-isoindol-2-yl)-piperidine-2,6-dione
methanesulfonic acid salt (0.37 g, 1 mmol) in DMF (2 mL) was added CDI (162 mg, 1
mmol). The mixture was stirred at RT overnight. 4-Phenylaniline (169 mg, 1 mmol) was
added to the mixture and the mixture was stirred at RT for 4 hrs. Then the temperature
was elevated to 40 °C and the mixture was stirred at this temperature for 8 hrs. The
mixture was cooled to RT. The suspension was filtered and the filtrate was added CH
3CN (5 mL) and the resulted suspension was filtered. The collected solid was recrystalized
from DMF to give the product as a white solid (30 mg, 15% yield): HPLC: Waters Symmetry
C
18, 5 µm, 3.9 x 150 mm, 1 mL/min, 240 nm, 50/50 CH
3CN/0.1% H
3PO
4, 2.78 min (95 %);
1H NMR (DMSO-
d6)
δ 2.02 (br. s., 1H, CHH), 2.28 - 2.47 (m, 1H, CHH), 2.60 (d,
J = 18.5 Hz, 1H, CHH), 2.82 - 3.04 (m, 1H, CHH), 4.23 - 4.59 (m, 4H, CH
2, CH
2), 5.11 (dd,
J = 4.9, 13.2 Hz, 1H, NCH), 6.73 - 6.83 (m, 1H, NH), 7.22 - 7.35 (m, 1H, Ar), 7.37
- 7.66 (m, 10H, Ar), 7.70 (d,
J = 7.9 Hz, 1H, Ar), 8.75 (s, 1H, NH), 10.98 (s, 1H, NH);
13C NMR (DMSO-
d6)
δ 22.51, 31.20, 42.80, 47.13, 51.58, 118.09, 121.89, 122.94, 125.99, 126.64, 126.85,
128.82, 130.30, 132.86, 139.93, 142.40, 144.81, 155.18, 167.95, 170.99, 172.85; LCMS
MH
+ = 423.
5.16 1-[2-(2,6-Dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-3-[3-(3-methyl-[1,2,4]oxadiazol-5-yl)-phenyl]-urea (Example disclosed for reference only)
[0228]

[0229] To a stirred suspension of 3-(3-methyl-[1,2,4]oxadiazol-5-yl)-phenylamine (0.22 g,
1.09 mmol) and 3-(5-aminomethyl-1-oxo-1,3-dihydro-isoindol-2-yl)-piperidine-2,6-dione
methanesulfonic acid salt (0.40 g, 1.09 mmol) in acetonitrile (5 mL) at RT was added
TEA (0.31 mL, 2.19 mmol). The mixture was stirred for 3 hrs, followed by addition
of 1N HCl (10 mL), which was stirred for 10 min. The mixture was purified by preparative
HPLC to give the product as a white solid (0.10 g, 19% yield): HPLC, Waters Symmetry
C
18, 5 µm, 3.9 x 150 mm, 1 ml/min, 240 nm, 30/70 CH
3CN/0.1% H
3PO
4, 4.37 min (99.3%); mp, 242-244 °C;
1H NMR (DMSO-
d6)
δ 1.92 - 2.07 (m, 1H, CH
H), 2.30 - 2.46 (m, 4H, C
H3, CH
H), 2.55 - 2.67 (m, 1H, CH
H), 2.82 - 3.02 (m, 1H, CH
H), 4.24 - 4.56 (m, 4H, C
H2, C
H2), 5.11 (dd,
J = 5.1, 13.2 Hz, 1H, NC
H), 6.91 (t,
J = 5.8 Hz, 1H, N
H), 7.39 - 7.80 (m, 6H, Ar
H), 8.37 (s, 1H, Ar
H), 9.05 (s, 1H, N
H), 10.98 (s, 1H, N
H);
13C NMR (DMSO-
d6)
δ 11.25, 22.49, 31.20, 42.83, 47.12, 51.58, 116.37, 120.24, 121.93, 121.96, 122.94,
123.77, 126.94, 129.90, 130.33, 141.41, 142.39, 144.68, 155.10, 167.61, 167.93, 170.99,
172.85, 174.86. LC/MS MH = 475; Anal. Calcd. For C
24H
22N
6O
5 +0.4 H
2O: C, 59.85; H, 4.77; N, 17.45; Found: C, 59.53; H, 4.68; N, 17.30.
5.17 1-(3-Aminophenyl)-3-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)urea (Example disclosed for reference only)
[0230]

[0231] Step 1: To a stirred mixture of 3-(5-aminomethyl-1-oxo-1,3-dihydro-isoindol-2-yl)-pipcridinc-2,6-dionc
hydrochloride (0.74 g, 2.00 mmol) and 3-nitropheny isocyanate (0.33 g, 2.00 mmol)
in acetonitrile (20 mL) was added TEA (0.56 mL, 4.00 mmol) at RT under nitrogen. After
12 hrs, the solid was filtered and purified by preparative HPLC (gradient: CH
3CN /H
2O: 15/85 for 5 min, to 100/0 in 10 min, 100/0 for 5 min). After evaporation of the
solvent, the residue was triturated in ether (20 mL) for 1 hr. The product was then
isolated by filtration and dried
in vacuo to give 1-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-3-(3-nitrophenyl)urea
as a yellow solid (0.34 g, 39% yield).
[0232] Step 2: To a solution of 1-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindofin-5-yl)methyl)-3-(3-nitrophenyl)urea
(0.33 g, 0.80 mmol) in DMF (80 mL) was added Pd-C (0.10 g, <10% weight). The reaction
mixture was hydrogenated with a Parr-shaker at 55 psi. After 12 hrs, the mixture was
filtered through a celite pad, which was washed with additional DMF (50 mL). The filtrate
was then evaporated and the residue was stirred in water (150 mL) for 3 hrs. The solid
was filtered, washed with additional water (50 mL), and dried. The crude product was
dissolved in DMF (50 mL), decolorizing carbon was added, and the reaction mixture
was stirred for 12 hrs. The mixture was then filtered through a celite pad, which
was washed with additional DMF (50 mL). The filtrate was then evaporated and the residue
was stirred in water (100 mL) for 3 hrs. The solid was filtered, washed with additional
water (50 mL) and dried
in vacuo to give the product as a pale yellow solid (0.24 g, 77% yield): HPLC: X-Terra RP
18, 3.9 X 150 mm, 5 µm, 1 mL/min, 240 nm: CH
3CN/0.1% (HCO
2)NH
4: 15/85: 7.95 min (95.27%); mp: 233-235 °C;
1H NMR (DMSO-
d6)
δ 1.78 - 2.08 (m, 1H, CHH), 2.38 (qd,
J = 4.5, 13.2 Hz, 1H, CHH), 2.59 (d,
J = 17.8 Hz, 1H, CHH), 2.79 - 3.06 (m, 1H, CHH), 4.18 - 4.36 (m, 1H, CHH), 4.36 - 4.60
(m, 3H, CHH, CH
2), 4.91 - 5.36 (m, 3H, NH
2, CH), 6.15 (ddd,
J = 0.9, 2.1, 7.9 Hz, 1H, Ar), 6.48 - 6.58 (m, 1H, Ar), 6.63 (t,
J = 6.0 Hz, 1H, NH), 6.76 (t,
J = 2.0 Hz, 1H, Ar), 6.85 (t,
J = 7.9 Hz, 1H, Ar), 7.44 (d,
J = 7.7 Hz, 1H, Ar), 7.51 (s, 1H, Ar), 7.69 (d,
J = 7.9 Hz, 1H, Ar), 8.32 (s, 1H, NH), 10.98 (s, 1H, NH);
13C NMR (DMSO-
d6)
δ 22.51, 31.20, 42.73, 47.12, 51.58, 103.78, 106.20, 107.79, 121.86, 122.91, 126.89,
128.89, 130.26, 140.92, 142.38, 144.99, 148.44, 155.16, 167.95, 170.99, 172.85; LCMS:
MH
+ = 408; Anal. Calcd. for C
21H
21N
5O
4: C, 61.91, H, 5.20, N, 17.19; Found: C, 62.40, H, 5.67, N, 15.59.
5.18 1-[2-(2,6-Dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-3-[3-(pyridin-2-yloxy)-phenyl]-urea (Example disclosed for reference only)
[0233]

[0234] 3-(5-aminomethyl-1-oxo-1,3-dihydro-isoindol-2-yl)-piperidine-2,6-dione methanesulfonate
(1.11 g, 3.0 mmol) and 1,1'-Carbonyldiimidazole (535 mg, 3.3 mmol) were suspended
in dry DMF (20 mL) and the mixture was stirred at rt for 24 h. While stirring, a portion
of the reaction mixture (6.7 mL, ∼1 mmol) was transferred to a vial containing 3-(Pyridin-2-yloxy)-phenylamine
(205 mg, 1.1 mmol). The resulting mixture was stirred at rt overnight and the reaction
progress was monitored by LCMS. After 48 h, additional 3-(Pyridin-2-yloxy)-phenylamine
(37 mg, 0.2 mmol) was transferred to the reaction mixture and stirring continued for
another 24 h. The reaction mixture was acidified with acidic acid and water. The volatiles
were removed in vacuo and the residue was dissolved in DMF and purified using C-18
preparatory HPLC to give 1-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-3-[3-(pyridin-2-yloxy)-phenyl]-urea
as a white solid (310 mg, 64% yield): HPLC:Waters Symmetry C
18, 5 µm, 3.9 x 150 mm, 1 ml/min, 240 nm, 30/70 CH
3CN/0.1% H
3PO
4, 4.81 min (98.6%); mp: 298-300 °C;
1H NMR (DMSO-d
6)
δ 1.84 - 2.06 (m, 1H, CHH), 2.19 - 2.44 (m, 1H, CHH), 2.54 - 2.68 (m, 1H, CHH), 2.79
- 3.05 (m, 1H, CHH), 4.08 - 4.60 (m, 4H, CH
2, CH
2), 5.10 (dd,
J = 5.1, 13.2 Hz, 1H, CH), 6.64 (dd,
J = 1.4, 8.0 Hz, 1H, Ar), 6.81 (t,
J = 5.9 Hz, 1H, NH), 6.99 (d,
J = 8.3 Hz, 1H, Ar), 7.07 - 7.18 (m, 2H, Ar), 7.24 (t,
J = 8.1 Hz, 1H, Ar), 7.33 (t,
J = 2.1 Hz, 1H, Ar), 7.43 (d,
J = 7.7 Hz, 1H, Ar), 7.50 (s, 1H, Ar), 7.68 (d,
J = 7.7 Hz, 1H, Ar), 7.78 - 7.97 (m, 1H, Ar), 8.16 (dd,
J = 1.5, 4.9 Hz, 1H, Ar), 8.82 (s, 1H, NH), 10.97 (s, 1H, NH);
13C NMR (DMSO-d
6) δ 22.49, 31.18, 42.73, 47.12, 51.56, 110.23, 111.51, 113.54, 113.68, 118.99, 121.85,
122.93, 126.86, 129.56, 130.29, 140.10, 141.79, 142.39, 144.77, 147.52, 154.37, 155.09,
163.03, 167.93, 170.99, 172.85; LCMS: MH = 486; Anal Calcd for C
26H
23N
5O
5 + 0.3 H
2O: C, 63.61; H, 4.85; N, 14.27. Found: C, 63.62; H, 4.62; N, 14.18.
5.19 1-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-3-(3-(piperidin-4-yloxy)phenyl)urea (Example disclosed for reference only)
[0235]

[0236] Using the procedure as described in Section 5.15, the product is prepared from 3-(piperidin-4-yloxy)aniline
and 3-(5-aminomethyl-1-oxo-1,3-dihydro-isoindol-2-yl)-piperidine-2,6-dione methanesulfonic
acid salt.
5.20 1-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-3-(3-hydroxy-4-methylphenyl)urea (Example disclosed for reference only)
[0237]

[0238] Step 1: Using the procedure as described in Section 5.15, 1-(3-(
tert-butyldimethylsilyloxy)-4-methylphenyl)-3-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)urea
is prepared from 3-(
tert-butyldimethylsilyloxy)-4-methylaniline and 3-(5-aminomethyl-1-oxo-1,3-dihydro-isoindol-2-yl)-piperidine-2,6-dione
methanesulfonic acid salt.
[0239] Step 2: A mixture of 1-(3-(
tert-butyldimethylsilyloxy)-4-methylphenyl)-3-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)urea
(0.54 g, 1.0 mmol) and cesium fluoride (0.15 g, 1.0 mmol) in DMF (10 mL) is heated
to 70 °C for 8 hrs. The mixture is cooled and diluted with water (10 mL). The solid
precipitate is filtered, rinsed with water (10 mL), and dried under vacuum to provide
the product.
5.21 5-(3-((2-(2,6-Dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)ureido)-2-methylphenyl
2-aminoacetate hydrochloride (Example disclosed for reference only)
[0240]

[0241] Step 1: Using the procedure as described in Section 5.15, 5-(3-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)ureido)-2-methylphenyl
2-(
tert-butoxycarbonylamino)acetate is prepared from 5-amino-2-methylphenyl 2-(
tert-butoxycarbonylamino)acetate and 3-(5-aminomethyl-1-oxo-1,3-dihydro-isoindol-2-yl)-piperidine-2,6-dione
methanesulfonic acid salt.
[0242] Step 2: To a mixture of 5-(3-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)ureido)-2-methylphenyl
2-(
tert-butoxycarbonylamino)acetate (0.58 g, 1.0 mmol) in DCM (50 mL) is added 2M HCl in
ether (1 mL), and the mixture is stirred for 24 hrs. The solid precipitate is filtered,
rinsed with DCM (10 mL), and dried under vacuum to provide the product.
5.22 5-(3-((2-(2,6-Dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)ureido)-2-methylphenyl
2-(piperazin-1-yl)acetate hydrochloride (Example disclosed for reference only)
[0243]

[0244] Step 1: Using the procedure as described in Section 5.15,
tert-butyl 4-(2-(5-(3-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)ureido)-2-methylphenoxy)-2-oxoethyl)piperazine-1-carboxylate
is prepared from
tert-butyl 4-(2-(5-amino-2-methylphenoxy)-2-oxoethyl)piperazine-1-carboxylate and 3-(5-aminomethyl-1-oxo-1,3-dihydro-isoindol-2-yl)-piperidine-2,6-dione
methanesulfonic acid salt.
[0245] Step 2: To a mixture of
tert-butyl 4-(2-(5-(3-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)ureido)-2-methylphenoxy)-2-oxoethyl)piperazine-1-carboxylate
(0.65 g, 1.0 mmol) in methylene chloride (50 mL) is added 2M HCl in ether (1 mL),
and the mixture will be stirred for 24 hrs. The solid precipitate is filtered, rinsed
with DCM (10 mL), and dried under vacuum to provide the product.
5.23 1-(3-(Aminomethyl)-4-methylphenyl)-3-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)urea (Example disclosed for reference only)
[0246]

[0247] Step 1: Using the procedure as described in Section 5.15, 1-(3-cyano-4-methylphenyl)-3-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)urea
is prepared from 5-amino-2-methylbenzonitrile and 3-(5-aminomethyl-1-oxo-1,3-dihydro-isoindol-2-yl)-piperidine-2,6-dione
methanesulfonic acid salt.
[0248] Step 2: A mixture of 1-(3-cyano-4-methylphenyl)-3-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)urea
(0.54 g, 1.0 mmol), platinum oxide (0.1 g), and 5-6M HCl in isopropanol (2 mL) in
acetic acid (15 mL) is hydrogenated under 50 psi hydrogen for 48 hrs. The mixture
is filtered through Celite, the filtrate is evaporated under vacuum, and the residue
is purified by preparative HPLC to provide the product.
5.24 1-(2-(Aminomethyl)-5-methylphenyl)-3-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)urea (Example disclosed for reference only)
[0249]

[0250] Step 1: Using the procedure as described in Section 5.15, 1-(2-cyano-5-methylphenyl)-3-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)urea
is prepared from 2-amino-4-methylbenzonitrile and 3-(5-aminomethyl-1-oxo-1,3-dihydro-isoindol-2-yl)-piperidine-2,6-dione
methanesulfonic acid salt.
[0251] Step 2: A mixture of 1-(2-cyano-5-methylphenyl)-3-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)urea
(0.54 g, 1.0 mmol), platinum oxide (0.1 g), and 5-6M HCl in isopropanol (2 mL) in
acetic acid (15 mL) is hydrogenated under 50 psi hydrogen for 48 hrs. The mixture
is filtered through Celite, the filtrate is evaporated under vacuum, and the residue
is purified by preparative HPLC to provide the product.
5.25 1-((2-(2,6-Dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-3-(4-(morpholinomethyl)phenyl)urea (Example disclosed for reference only)
[0252]

[0253] Using the procedure as described in Section 5.15, the product is prepared from 4-(morpholinomethyl)aniline
and 3-(5-aminomethyl-1-oxo-1,3-dihydro-isoindol-2-yl)-piperidine-2,6-dione methanesulfonic
acid salt.
5.26 1-((2-(2,6-Dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-3-(3-((4-methylpiperazin-1-yl)methyl)phenyl)urea (Example disclosed for reference only)
[0254]

[0255] Using the procedure as described in Section 5.15, the product is prepared from 3-((4-methylpiperazin-1-yl)methyl)aniline
and 3-(5-aminomethyl-1-oxo-1,3-dihydro-isoindol-2-yl)-piperidine-2,6-dione methanesulfonic
acid salt.
5.27 Isoindolin compounds (Example disclosed for reference only)
[0256]

[0257] The isoindolin compounds shown above are made using the procedure as described in
Section 5.15.
5.28 1-[2-(2,6-Dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-3-[4-(2-pyridin-4-yl-ethyl)-phenyl]-urea
hydrochloric acid salt (Example disclosed for reference only)
[0258]

[0259] Step 1: Preparation of 4-[2-(4-nitro-phenyl)-vinyl]-pyridine To a CH
3CN solution of 1-iodo-4-nitrobenzene (498 mg, 2 mmol) and 4-vinylpyridinc (262 mg,
2.5 mmol) was added TEA (350 µL, 2.5 mmol) and Pd(OAc)
2 (0.45 mg, 0.2 mmol). The mixture was heated to 100°C for 48 hr in a capped Pyrex
tube. It was then cooled to room temperature, quenched with 1N HCl (20 mL) and concentrated
under reduced pressure. The mixture was filtered and the solid was taken up in EtOAc
(30 mL) and washed with 1N NaOH (30 mL). The organic layer was concentrated, dried
over Na
2SO
4 and concentrated to give 4-[2-(4-nitro-phenyl)-vinyl]-pyridine as yellow solid (160
mg, 37%).
[0260] Step 2: Preparation of 4-(2-pyridin-4-yl-ethyl)-phenylamine To the EtOAc solution (30 mL)
of 4-[2-(4-nitro-phenyl)-vinyl]-pyridine (160 mg, 0.71 mmol) was added palladium on
carbon (0.1 g, 50% wet). The suspension was hydrogenated at 50 psi of hydrogen for
2 hours. The mixture was filtered over a celite pad. The filtrate was concentrate
to give 4-(2-pyridin-4-yl-ethyl)-phenylamine as a yellow solid (140 mg, 100%).
[0261] Step 3: Preparation of 1-[2-(2,6-Dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-3-[4-(2-pyridin-4-yl-ethyl)-phenyl]-urea
hydrochloric acid salt To a suspension of 3-(5-aminomethyl-1-oxo-1,3-dihydro-isoindol-2-yl)-piperidine-2,6-dione
methanesulfonic acid salt (0.37 g, 1 mmol) in DMF (2 mL) was added carbonyl diimidazole
(162 mg, 1 mmol). The mixture was stirred at room temperature overnight. 4-Pyridin-4-ylethyl-phenylamine
(140 mg, 0.7 mmol) was added to the mixture and the mixture was stirred at 40 °C overnight
then 80°C for 1.5 hours. The mixture was cooled to room temperature, added water (5
mL) and stirred for 10 min. The suspension was filtered and the filtrate was concentrated
and purified on prep-HPLC. The resulted solid after purification was stirred with
1N HCl, filtered and the filtrate was concentrated to give1-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-3-[4-(2-pyridin-4-yl-ethyl)-phenyl]-urea
hydrochloric acid salt as an off-white solid (30 mg, 8% yield). HPLC: Waters Symmetry
C
18, 5µm, 3.9 x 150 mm, 1 mL/min, 240 nm, 5% grad 95% in 5 min, CH
3CN/0.1% H
3PO
4,: t
R =4.70 min (92%); mp: > 400 °C;
1H NMR (DMSO-d
6) δ 1.89 - 2.06 (m, 1H, CHH), 2.37 (br. s., 1H, CHH), 2.55 - 2.67 (m, 1H, CHH), 2.82
- 3.01 (m, 3H, CH
2, CHH), 3.12 (d,
J = 7.7 Hz, 2H, CH
2), 4.20 - 4.53 (m, 5H, CH
2, CH
2), 5.11 (dd,
J = 4.8, 13.3 Hz, 1H, NCH), 6.89 (br. s., 1H, NH), 7.07 (d,
J = 8.3 Hz, 2H, Ar), 7.32 (d,
J = 8.3 Hz, 2H, Ar), 7.44 (d,
J = 7.9 Hz, 1H, Ar), 7.51 (s, 1H, Ar), 7.69 (d,
J = 7.7 Hz, 1H, Ar), 7.83 (d,
J = 5.5 Hz, 2H, Ar), 8.71 - 8.78 (m, 2H, Ar), 8.80 (s, 1H, NH), 10.98 (s, 1H, NH).
13C NMR (DMSO-d
6) δ 22.44,31.13, 34.25, 36.52, 42.64, 47.05, 51.49, 117.59, 121.78, 122.84, 126.56,
126.78, 128.49, 130.19, 132.39, 138.59, 142.12, 142.28, 144.86, 155.24, 167.86, 170.91,
172.78; LCMS MH = 498; Anal Calcd for C
28H
27N
5O
4+ 1.55 HCl + 0.75H
2O + 0.15 CH
3CN C, 56.78; H, 5.29; N, 11.64; Cl, 8.87; Found: C, 56.47; H, 5.15; N, 11.99; Cl,
8.85.
5.29 Isoindolin compounds (Example disclosed for reference only)
[0262]

[0263] These isoindolin compounds shown above are made using the procedure as described
in Section 5.15.
5.30 N"-(3-Chloro-4-methyl-phenyl)-N'-[2-(2,6-dioxo-niperidin-3-yl)-1-oxo-2, 3-dihydro-1H-isoindol-5-ylmethyl]-N-cyano-guanidine (Example disclosed for reference only)
[0264]

[0265] Step 1: Preparation of (3-chloro-4-methyl-phenyl)-carbamic acid phenyl ester. 2-chloro-4-amino
toluene (282 mg, 2 mmol) was dissolved in THF (10 mL). The mixture was added sodium
hydride (128 mg, 3.2 mmol) and stirred at room temperature for 15 minutes. Dipehnyl
N-cyano-carbonimidate (715 mg, 3.0 mmol) was added and the mixture was heated to reflux
for 4 hours. The reaction mixture was cooled to room temperature, quenched by saturated
NH
4Cl (10 mL), filtered and the solid was dried in oven to give (3-chloro-4-methyl-phenyl)-carbamic
acid phenyl ester as solid (0.5 g, 87%).
[0266] Step 2: Preparation of N"-(3-Chloro-4-methyl-phenyl)-N'-[2- (2,6-dioxo-piperidin-3-yl)-1-oxo-2,
3-dihydro-1H-isoindol-5-ylmethyl]-N-cyano-guanidine To a suspension of 3-(5-aminomethyl-1-oxo-1,3-dihydro-isoindol-2-yl)-piperidine-2,6-dione
methanesulfonic acid salt (0.18 g, 0.5 mmol) in DMF (2 mL), was added (3-chloro-4-methyl-phenyl)-carbamic
acid phenyl ester (143 mg, 0.5 mmol) and DIPEA (83 µL, 0.5 mmol). The mixture was
stirred at 100°C for 4 hours. The mixture was concentrated under reduced pressure
to remove DMF, purified on silica gel column eluted using methanol and methylene chloride
to give N"-(3-Chloro-4-methyl-phenyl)-N'-[2- (2,6-dioxo-piperidin-3-yl)-1-oxo-2, 3-dihydro-1H-isoindol-5-ylmethyl]-N-cyano-guanidine
as a white solid (40 mg,17% yield). HPLC: Waters Symmetry C
18, 5µm, 3.9 x 150 mm, 1 mL/min, 240 nm, 50/50, CH
3CN/0.1% H
3PO
4,: t
R =3.06 min (96.5%); mp: 325-327 °C;
1H NMR (DMSO-d
6) δ 1.93 - 2.09 (m, 1H, CHH), 2.25 - 2.32 (m, 3H, CH
3), 2.33 - 2.45 (m, 1H, CHH), 2.55 - 2.67 (m, 1H, CHH), 2.82 - 3.01 (m, 1H, CHH), 4.27
- 4.57 (m, 4H, CH
2,CH
2), 5.11 (dd,
J = 5.1, 13.2 Hz, 1H, NCH), 7.13 (dd,
J = 2.3, 8.1 Hz, 1H, Ar), 7.32 (d,
J = 1.5 Hz, 2H, Ar), 7.43 (d,
J = 7.7 Hz, 1H, Ar), 7.51 (s, 1H, Ar), 7.71 (d,
J = 7.7 Hz, 1H, Ar), 7.85 (s, 1H, NH), 9.20 (s, 1H, NH), 10.99 (s, 1H, NH).
13C NMR (DMSO-d
6) δ 18.98, 22.49, 31.20, 44.70, 47.15, 51.58, 116.88, 121.98, 122.94, 124.33, 126.97,
130.52, 131.41, 132.08, 133.10, 136.46, 142.36, 142.97, 158.15, 167.86, 170.98, 172.86.;
LC-MS: 465; Anal Calcd for C
23H
21ClN
6O
3 + 0.6 H
2O + 0.2 EtOAc: C, 57.94; H, 4.86; N, 17.03;Found: C, 57.66; H, 4.81; N, 17.10.
5.31 1-[2-(2,6-Dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-3-[4-(1H-imidazol-2-yl)-phenyl]-urea (Example disclosed for reference only)
[0267]

[0268] A mixture of 3-(5-aminomethyl-1-oxo-1,3-dihydro-isoindol-2-yl)-piperidine-2,6-dione
methanesulfonic acid salt (0.40 g, 1.1 mmol) and CDI (0.19 g, 1.2 mmol) in DMF (5
mL) was stirred at room temperature for 18 hours. To the mixture was added 4-(1H-imidazol-2-yl)-phenylamine
(0.17 g, 1.1 mmol) at room temperature, and the mixture was stirred at 60 C for 1
d. To the mixture was added water (25 mL) and ethyl acetate (20 mL). The mixture was
stirred at room temperature for 2 h. The suspension was filtered and the solid was
washed with water (20 mL), ethyl acetate (20 mL) and water (20 mL) to give a solid.
The solid was purified with Prep HPLC to give 1-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-3-[4-(1H-imidazol-2-yl)-phenyl]-urea
as a white solid (150 mg, 30% yield): HPLC: Waters Symmetry C
18, 5µm, 3.9 x 150 mm, 1 mL/min, 240 nm, 5/95 grad 95/5 in 5 min CH
3CN/0.1% H
3PO
4, 4.41 min (95.9 %); mp: 190-192°C;
1H NMR (DMSO-d
6) δ 1.91 - 2.08 (m, 1H, CHH), 2.27 - 2.47 (m, 1H, CHH), 2.54 - 2.66 (m, 1H, CHH),
2.81 - 3.01 (m, 1H, CHH), 4.31 (d,
J = 17.4 Hz, 1H, CHH), 4.38 - 4.52 (m, 3H, CHH, CH
2), 5.11 (dd,
J = 5.0, 13.3 Hz, 1H, NCH), 6.78 (t,
J = 6.0 Hz, 1H, NH), 7.33 - 7.76 (m, 9H, Ar), 8.17 (s, 1H, HCOOH), 8.67 (s, 1H, NH),
10.99 (s, 1H, NH);
13C NMR (DMSO-d
6) δ 22.41, 31.10, 42.69, 47.02, 51.49, 114.17 (br), 117.80, 121.79, 122.84, 124.55,
126.81, 126.98, 130.19, 135.46, 138.55, 142.31, 144.83, 155.14, 163.19 (HCOOH), 167.88,
170.92, 172.77; LCMS MH = 459; Anal. Calcd for C
24H
22N
6O
4 + 2 H
2O + 0.7 HCOOH + 0.3 DMF: C, 56.04; H, 5.42; N, 16.08; Found: C, 55.84; H, 5.34; N,
16.11.
5.32 1-[3-(1H-Benzoimidazol-2-yl)-4-chloro-phenyl]-3-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-urea (Example disclosed for reference only)
[0269]

[0270] A mixture of 3-(5-aminomethyl-1-oxo-1,3-dihydro-isoindol-2-yl)-piperidine-2,6-dione
methanesulfonic acid salt (0.40 g, 1.1 mmol) and CDI (0.21 g, 1.3 mmol) in DMF (5
mL) was stirred at room temperature for 18h. To the mixture was added 3-(1H-benzoimidazol-2-yl)-4-chloro-phenylamine
(0.32 g, 1.3 mmol) at room temperature, and the mixture was stirred at 100 °C for
18 hours. To the mixture was added water (25 mL) and ether (20 mL). The mixture was
stirred at room temperature for 2 h. The solvent was decanted. The solid was purified
with Prep HPLC to give 1-[3-(1H-benzoimidazol-2-yl)-4-chloro-phenyl]-3-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-urea
as a white solid (130 mg, 22% yield): HPLC: Waters Symmetry C
18, 5µm, 3.9 x 150 mm, 1 mL/min, 240 nm, 20/80 CH
3CN/0.1% H
3PO
4, 4.44 min (99.2 %); mp: 275-277 °C;
1H NMR (DMSO-d
6) δ 1.90 - 2.10 (m, 1H, CHH), 2.23 - 2.46 (m, 1H, CHH), 2.54 - 2.68 (m, 1H, CHH),
2.80 - 3.02 (m, 1H, CHH), 4.31 (d,
J = 17.4 Hz, 1H, CHH), 4.37 - 4.51 (m, 3H, CHH, CH
2), 5.11 (dd,
J = 5.0, 13.3 Hz, 1H, NCH), 6.89 (t,
J = 5.9 Hz, 1H, NH), 7.23 (d,
J = 4.5 Hz, 2H, Ar), 7.40 - 7.76 (m, 7H, Ar), 8.07 (d,
J = 2.5 Hz, 1H, Ar), 9.02 (s, 1H, NH), 10.98 (s, 1H, NH), 12.64 (br. s., 1H, NH);
13C NMR (DMSO-d
6) δ 22.39, 31.10, 42.75, 47.02, 51.48, 111.62, 118.88, 120.11, 120.62, 121.55, 121.83,
122.56, 122.85, 126.85, 129.79, 130.23, 130.35, 134.56, 139.56, 142.29, 143.00, 144.58,
149.12, 154.96, 162.96, 167.86, 170.91, 172.76; LCMS MH = 543, 545; Anal. Calcd for
C
28H
23N
6O
4Cl + 3 H
2O: C, 56.33; H, 4.90; N, 14.08; Found: C, 56.73; H, 4.67; N, 14.09.
5.33 N-(3-Chloro-4-methyl-phenyl)-N'-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-guanidine
Hydrochloride (Example disclosed for reference only)
[0271]

Step 1: 3-Chloro-4-methyl-phenyl)-thiourea
[0272] To a solution of 2-chloro-4-isothiocyanato-1-methyl-benzene (2 gm, 10.89 mmol) in
dry acetonitrile (10 mL) was added a solution of NH
3 in MeOH (2 N, 6 mL) and the mixture was stirred at rt for 2 h. Water was added to
the reaction mixture causing the a precipitate to form. The reaction slurry was concentrated
in vacuo to ∼1/4 the volume and the white solid was collected by filtration and dried
in a vacuum oven to give 2.1 gm (96% yield) of 3-chloro-4-methyl-phenyl)-thiourea
as a white solid that was used without further purification. LCMS: MH = 201.
Step 2: 1-(3-Chloro-4-methyl-phenyl)-2-methyl-isothiourea Hydroiodide
[0273] To a slurry of 3-chloro-4-methyl-phenyl)-thiourea (2.1 gm, 10.46 mmol) in MeOH (15
mL) was added methyl iodide (2.5 mL, 40 mmol) and the mixture was stirred at rt for
4 h. The reaction mixture was concentrated to dryness and to the residue, a small
portion of dichloromethane was added and the volatiles removed in vacuo. This process
was repeated twice more to provide a tan foam which was dried in a vacuum oven overnight
to give 3.5 gm (98% yield) of crude (2x). 1-(3-chloro-4-methylphenyl)-2-methyl-isothiourea
hydroiodide. This material was found to be highly hygroscopic and was stored in a
desiccator for further use. LCMS: MH = 215.
Step 3: N-(3-Chloro-4-methyl-phenyl)-N'-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-guanidine
Hydrochloride
[0274] In a microwave vial fitted with a stir bar, 1-(3-chloro-4-methyl-phenyl)-2-methyl-isothiourea
hydroiodide (597 mg, 1.74 mmol), 3-(5-aminomethyl-1-oxo-1,3-dihydro-isoindol-2-yl)-piperidine-2,6-dione
methanesulfonate (560 mg, 1.52mmol), and DIEA (1.1 mL, 6.06 mmol) were combined in
anhydrous DMF (5 mL). The vial was sealed and irradiated in a microwave for 30 min
at 120 °C. To the reaction mixture was added an additional 100 mg of 3-(5-aminomethyl-1-oxo-1,3-dihydro-isoindol-2-yl)-piperidine-2,6-dione
methanesulfonate and the reaction was irradiated for 30 min at 120 °C. This process
was repeated twice more to consume all of the 1-(3-chloro-4-methyl-phenyl)-2-methyl-isothiourea
hydroiodide starting material as it was found co-elute with the desired product on
LCMS. The reaction mixture was acidified with acetic acid and the volatiles were evaporated
in vacuo. The residue was dissolved in minimal DMF, filtered, and purified using C-18
preparatory HPLC. To the combined fractions containing the desired product, 1 N HCl
was added and the solvents removed in vacuo. The obtained white solid was dissolved
in minimal water and lyophilized to give 180 mg (27% yield) of N-(3-chloro-4-methyl-phenyl)-N'-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-guanidine
hydrochloride: HPLC:Waters Symmetry C
18, 5 µm, 3.9 x 150 mm, 1 ml/min, 240 nm, 20/80 CH
3CN/0.1% H
3PO
4, 3.73 min (97.4%); mp: >400 °C;
1H NMR (DMSO-d
6) δ 1.91 - 2.09 (m, 1H, CHH), 2.33 (s, 3H, CH
3), 2.41 (dd,
J = 4.3, 13.2 Hz, 1H, CHH), 2.54 - 2.68 (m, 1H, CHH), 2.80 - 3.05 (m, 1H, CHH), 4.21
- 4.56 (m, 2H, CH
2), 4.66 (d,
J = 5.7 Hz, 2H, CH
2), 5.13 (dd,
J = 4.9, 13.2 Hz, 1H, CH), 7.14 (dd,
J = 1.9, 8.1 Hz, 1H, Ar), 7.34 (d,
J = 1.9 Hz, 1H, Ar), 7.42 (d,
J = 8.1 Hz, 1H, Ar), 7.51 (d,
J = 7.9 Hz, 1H, Ar), 7.61 (s, 1H, Ar), 7.76 (d,
J = 7.7 Hz, 1H, Ar), 7.98 (br. s., 1H, NH), 8.62 (br. s., 1H, NH), 10.16 (s, 1H, NH),
11.00 (s, 1H, NH);
13C NMR (DMSO-d
6) δ 21.04, 24.19, 32.95, 46.33, 49.85, 54.03, 124.13, 125.48, 125.79, 127.28, 129.17,
132.57, 134.20, 135.82, 135.95, 136.64, 143.17, 144.68, 156.84, 170.94, 173.00, 175.82;
LCMS: MH = 440, 442; Anal Calcd for C
22H
23Cl
2N
5O
3 + 2.3 H
2O + 1.9 HCl + 0.2 HCOOH: C, 44.72; H, 5.05; N, 11.74; Cl, 23.19; Found: C, 44.55;
H, 4.71; N, 11.48; Cl, 22.92.
5.34 1-((2-(2,6-Dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-3-(3-hydroxy-4-methylphenyl)urea (Example disclosed for reference only)
[0275]

[0276] Step 1: A mixture of 2-methyl-5-nitrophenol (3.0 g, 19.6 mmol) TBS-Cl (3.0 g, 19.6 mmol),
and TEA (2.2 g, 21.6 mmol) in DMF (30 mL) stirred at ambient temperature for 16 h.
The mixture was evaporated under vacuum. The residue was dissolved in ethyl acetate
(100 mL) and washed with water (3 x 100 mL) and evaporated under vacuum. The residue
was chromatographed in a hexanes-ethyl acetate gradient, providing 4.7 g of tert-butyldimethyl(2-methyl-5-nitrophenoxy)silane,
in 91% yield;
1H NMR (DMSO-d
6) δ 0.27 (s, 6H), 1.01 (s, 9H), 2.27 (s, 3H), 7.47 (d,
J = 8.5 Hz, 1H), 7.52 (d,
J = 2.3 Hz, 1H), 7.78 (dd,
J = 2.3, 8.3 Hz, 1H).
[0277] Step 2: A mixture of the product from Step 1 (4.7 g, 17.6 mmol) and 10% Pd-C (1.0 g, 50%
wet) in ethyl acetate was hydrogenated under 50 psi hydrogen for 16 h. The mixture
was then filtered through Celite and the filtrate was evaporated under vacuum, providing
3.9 g of -(tert-butyldimethylsilyloxy)-4-methylaniline, in 94% yield;
1H NMR (DMSO-d
6) δ 0.17 (s, 6H), 0.97 (s, 9H), 1.96 (s, 3H), 4.81 (s, 2H), 6.00 - 6.21 (m, 2H), 6.74
(d,
J = 7.9 Hz, 1H).
[0278] Step 3: A mixture of 3-(5-(aminomethyl)-1-oxoisoindolin-2-yl)piperidine-2,6-dione methanesulfonate
(0.74 g, 2.0 mmol) and CDI (0.32 g, 2.0 mmol) in DMF (30 mL) was stirred at room temperature
for 3 h, and then the product from Step 2 (0.47 g, 2.0 mmol) was added. The mixture
was heated to 70 °C for 16 h. Then, the mixture was cooled to room temperature and
quenched with 1N HCl (30 mL). The resulting mixture was evaporated under vacuum and
the residue was purified by preparative HPLC using an acetonitrile-water gradient,
and providing 0.38 g of 1-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-3-(3-hydroxy-4-methylphenyl)urea
as a white solid, in 46% yield; HPLC:Waters Symmetry C
18, 5 µm, 3.9 x 150 mm, 1 ml/min, 240 nm, 25/75 CH
3CN/0.1% H
3PO
4, 4.80 min (100.00%); mp 273-275 °C;
1H NMR (DMSO-d
6) δ 2.02 (s, 4H), 2.27 - 2.47 (m, 1H), 2.63 (m., 1H), 2.80 - 3.04 (m, 1H), 4.17 -
4.59 (m, 4H), 5.11 (dd,
J = 4.7, 13.0 Hz, 1H), 6.47 - 6.72 (m, 2H), 6.86 (d,
J = 7.9 Hz, 1H), 7.04 (s, 1H), 7.44 (d,
J = 7.7 Hz, 1H), 7.51 (s, 1H), 7.69 (d,
J = 7.7 Hz, 1H), 8.39 (s, 1H), 9.13 (s, 1H), 10.98 (s, 1H);
13C NMR (DMSO-d
6) δ 15.35, 22.49, 31.20, 42.74, 47.12, 51.58, 104.76, 108.45, 116.48, 121.86, 122.91,
126.91, 130.19, 130.26, 138.92, 142.36, 144.96, 155.18, 155.29, 167.95, 170.99, 172.85;
LCMS MH = 423; Anal. Calcd for C
22H
22N
4O
5: C, 62.55; H, 5.25; N, 13.26; Found: C, 62.27; H, 5.15; N, 13.18.
5.35 1-((2-(2,6-Dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-3-(4-hydroxy-3-methylphenyl)urea (Example disclosed for reference only)
[0279]

[0280] Step 1: A mixture of 2-methyl-4-nitrophenol (3.0 g, 19.6 mmol), TBS-Cl (3.0 g, 19.6 mmol),
and TEA (2.2 g, 21.6 mmol) in DMF (30 mL) stirred at ambient temperature for 16 h.
The mixture was partitioned between 10% aqueous sodium bicarbonate solution (100 mL)
and ethyl acetate (100 mL), and the aqueous layer was extracted with ethyl acetate
(100 mL). The combined organic layers were washed with 10% aqueous sodium bicarbonate
(3 x 100 mL) and water (100 mL), and evaporated under vacuum. The residue was chromatographed
in hexanes-ethyl acetate gradient, providing 3.9 g of tert-butyldimethyl(2-methyl-4-nitrophenoxy)silane,
in 75% yield;
1H NMR (DMSO-d
6) δ 0.29 (s, 6H), 1.00 (s, 9H), 2.24 (s, 3H), 7.03 (d,
J = 8.9 Hz, 1H), 8.02 (dd,
J = 2.8, 8.9 Hz, 1H), 8.12 (d,
J = 2.5 Hz, 1H).
[0281] Step 2: A mixture of the product from step 1 (3.9 g, 14.6 mmol) and 10% Pd-C (0.5 g, 50%
wet) in ethyl acetate was hydrogenated under 50 psi hydrogen for 18 h. The mixture
was filtered through Celite and the filtrate was evaporated, providing 3.2 g of 4-(tert-butyldimethylsilyloxy)-3-methylaniline,
in 93% yield.
[0282] Step 3: A mixture of 3-(5-(aminomethyl)-1-oxoisoindolin-2-yl)piperidine-2,6-dione methanesulfonate
(0.74 g, 2.0 mmol) and CDI (0.32 g, 2.0 mmol) in DMF (30 mL) was stirred at room temperature
for 3 h, and then the product from Step 2 (0.47 g, 2.0 mmol) was added. The mixture
was heated to 70 °C for 30 h. Then, the mixture was cooled to room temperature and
quenched with 1N HCl (30 mL), resulting in solid precipitate. This solid was purified
by chromotagraphy on a silica gel column, using a methylene chloride-methanol gradient,
and providing 0.59 g of 1-(4-(tert-butyldimethylsilyloxy)-3-methylphenyl)-3-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)urea
as an off-white solid, in 55% yield;
1H NMR (DMSO-d
6) δ 0.16 (s, 6H), 0.97 (s, 9H), 1.88 - 2.06 (m, 1H), 2.10 (s, 3H), 2.25 - 2.47 (m,
1H), 2.53 - 2.70 (m, 1H), 2.80 - 3.02 (m, 1H), 4.31 (d,
J = 17.4 Hz, 1H), 4.3 6 - 4. 52 (m, 3H), 5.11 (dd,
J = 4.9, 13.2 Hz, 1H), 6.51 - 6.74 (m, 2H), 7.08 (dd,
J = 2.5, 8.6 Hz, 1H), 7.18 (d,
J = 2.3 Hz, 1H), 7.43 (d,
J = 7.7 Hz, 1H), 7.50 (s, 1H), 7.69 (d,
J = 7.7 Hz, 1H), 8.35 (s, 1H), 10.98 (s, 1H).
[0283] Step 4: To a solution of the product from Step 3 (0.5 g, 0.9 mmol) in methylene chloride
(20 mL) was added 2N HCl in ether (2 mL). The mixture stirred for 16 h at ambient
temperature. The precipitated product was isolted by filtration, and was rinsed with
methylene chloride (20 mL) and dried under vacuum, providing 0.4 g of 1-((2-(2,6-dioxopiperidin-3-yl)-1
oxoisoindolin-5-yl)methyl)-3-(4-hydroxy-3-methylphenyl)urea as an off-white solid,
in quantitaive yield; HPLC:Waters Symmetry C
18, 5 µm, 3.9 x 150 mm, 1 ml/min, 240 nm, 30/70 CH
3CN/0.1% H
3PO
4, 1.88 min (96.94%); mp 210-212 °C;
1H NMR (DMSO-d
6) δ 1.94 - 2.18 (m, 4H), 2.30 - 2.51 (m, 1H), 2.58 - 2.72 (m, 1H), 2.87 - 3.06 (m,
1H), 4.25 - 4.56 (m, 4H), 5.15 (dd,
J = 5.0, 13.3 Hz, 1H), 6.67 (d,
J = 8.5 Hz, 1H), 7.02 (dd,
J = 2.5, 8.6 Hz, 1H), 7.12 (d,
J = 2.3 Hz, 1H), 7.47 (d,
J = 7.7 Hz, 1H), 7.54 (s, 1H), 7.73 (d,
J = 7.7 Hz, 1H), 8.32 (br. s., 1H), 11.02 (s, 1H);
13C NMR (DMSO-d
6) δ 16.15, 22.51, 31.20, 42.77, 47.12, 51.56, 114.47, 117.11, 121.32, 121.83, 122.88,
123.61, 126.86, 130.22, 131.72, 142.35, 145.17, 150.10, 155.58, 170.99, 172.85; LCMS
MH = 423; Anal. Calcd for C
22H
22N
4O
5 + 0.25 CH
2Cl
2: C, 60.23; H, 5.11; N, 12.63; Found: C, 60.14; H, 5.43; N, 12.54.
5.36 1-(4-tert-Butyl-evelohexyl)-3-12-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-urea (Example disclosed for reference only)
[0284]

[0285] To a stirred suspension of 4-
tert-butyl-cyclohexylamine (0.20 g, 1.28 mmol) in DMF (5 mL) at 40 °C was added CDI (0.23
g, 1.40 mmol). The mixture was stirred for 15 min, followed by addition of 3-(5-aminomethyl-1-oxo-1,3-dihydro-isoindol-2-yl)-piperidine-2,6-dione
methanesulfonic acid salt (0.47 g, 1.28 mmol). Heating was stopped after 1.5 hrs and
the mixture was stirred at RT overnight. Solvent was evaporated and the residue was
purified by preparative HPLC to give the product as a white solid (0.076 g, 13% yield):
HPLC, Waters Symmetry C18, 5 µm, 3.9 x 150 mm, 1 ml/min, 240 nm, 35/65 CH3CN/0.1% H3PO4, 13.37 min (44.7%), 14.68 min (54.3%); mp, 182-184 °C; 1H NMR (DMSO-d6) δ 0.83 (s, 9H, CH3, CH3, CH3), 0.90 - 1.21 (m, 4H, CH2, CH2), 1.30 - 1.58 (m, 2H, CHH, CHH), 1.61 - 1.78 (m, 2H, CHH, CHH), 1.87 (br. s., 1H, CHH), 1.95 - 2.06 (m, 1H, CHH), 2.28 - 2.47 (m, 1H, CHH), 2.63 (br. s., 1H, CHH), 2.83 - 3.00 (m, 1H, CHH), 3.78 & 3.27 (2s, 1H, CH), 4.23 - 4.50 (m, 4H, CH2, CH2), 5.11 (dd, J = 5.0, 13.1 Hz, 1H, NCH), 5.79 - 6.12 (m, 1H, NH), 6.25 - 6.44 (m, 1H, NH), 7.38 (t, J = 6.8 Hz, 1H, ArH), 7.45 (d, J = 6.2 Hz, 1H, ArH), 7.67 (dd, J = 4.1, 7.6 Hz, 1H, ArH), 10.99 (s, 1H, NH); (Note: 1H NMR showed about 55% to 45% isomer ratio); 13C NMR (DMSO-d6) δ 21.33, 22.49, 25.95, 27.38, 27.45, 31.02, 31.20, 32.06, 32.26, 33.76, 42.87, 43.43,
46.81, 47.10, 47.42, 48.69, 51.56, 121.76, 121.88, 122.83, 122.90, 126.81, 126.88,
130.11, 130.19, 142.29, 142.35, 145.36, 145.52, 157.35, 167.98, 170.99, 172.85. LC/MS
MH+ = 455; Anal. Calcd. For C25H34N4O4 +0.7 H2O: C, 64.27; H, 7.64; N, 11.99; Found: C, 63.98; H, 7.98; N, 11.92.
5.37 1-[2-(2,6-Dioxo-nioeridin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-3-(4-methyl-cyclohexyl)-urea (Example disclosed for reference only)
[0286]

[0287] To a stirred suspension of 4-methyl-cyclohexylamine (mixture of
cis/
trans isomers, 0.21 g, 1.86 mmol) in DMF (10 mL) at 40 °C was added CDI (0.33 g, 2.04 mmol).
The mixture was stirred for 15 min, followed by addition of 3-(5-aminomethyl-1-oxo-1,3-dihydro-isoindol-2-yl)-pipcridinc-2,6-dionc
methanesulfonic acid salt (0.69 g, 1.86 mmol). Heating was stopped after 1.5 hrs and
the mixture was stirred at RT overnight. Solvent was evaporated and the residue was
purified by preparative HPLC to give the product as a white solid (0.14 g, 18% yield):
HPLC, Waters Symmetry C
18, 5 µm, 3.9 x 150 mm, 1 ml/min, 240 nm, 25/75 CH
3CN/0.1% H
3PO
4, 11.33 min (59.1%), 12.41 min (39.7%); mp, 223-225 °C;
1H NMR (DMSO-
d6)
δ 0.81 - 0.93 (m, 3H, C
H3), 0.95 - 1.35 (m, 3H, CH
H, CH
H, CH
H), 1.37 - 1.57 (m, 4H, CH
H, CH
H, CH
H, C
H), 1.58 - 1.70 (m, 1H, CH
H), 1.73 - 1.88 (m, 1H, CH
H), 1.92 - 2.07 (m, 1H, CH
H), 2.30 - 2.47 (m, 1H, CH
H), 2.54 - 2.67 (m, 1H, CH
H), 2.83 - 3.00 (m, 1H, CH
H), 3.20-3.75 (m, 1 H, C
H), 4.21 - 4.51 (m, 4H, C
H2, C
H2), 5.11 (dd,
J = 5.0, 13.1 Hz, 1H, NC
H), 5.78 - 6.11 (m, 1H, N
H), 6.24 - 6.40 (m, 1H, N
H), 7.34 - 7.42 (m, 1H, Ar
H), 7.42 - 7.50 (m, 1H, Ar
H), 7.63 - 7.72 (m, 1H, Ar
H), 10.98 (s, 1H, N
H); (Note:
1H NMR showed about 60% to 40% isomer ratio);
13C NMR (DMSO-
d6)
δ 21.47, 22.17, 22.49, 29.42, 29.77, 30.42, 31.20, 31.50, 33.28, 33.73, 42.86, 44.54,
47.09, 48.37, 51.56, 121.78, 122.87, 126.81, 130.16, 142.33, 145.44, 145.52, 157.33,
167.98, 170.99, 172.85; LC/MS MH
+ = 413; Anal. Calcd. For C
22H
28N
4O
4 +0.4 H
2O: C, 63.23; H, 6.90; N, 13.41; Found: C, 62.93; H, 6.92; N, 13.09.
5.38 1-(3-Diethylamino-propyl)-3-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-urea,
formic acid salt
[0288]

[0289] To the solution of
para-nitrophenyl chloroformatc (202 mg, 1 mmol) in CH
3CN (5 mL) was added dropwise at 0 °C a solution of
N,N-diethyl-propane-1,3-diamine (130 mg, 1 mmol) and DIPEA (0.082 mL, 1 mmol) in CH
3CN (5 mL). The mixture was stirred at 0 °C for 10 min. 3-(5-Aminomethyl-1-oxo-1,3-dihydro-isoindol-2-yl)-piperidine-2,6-dione
methanesulfonic acid salt (0.37 g, 1 mmol) was added, followed by additional DIPEA
(0.082 mL, 1 mmol). The mixture was allowed to warm to RT and stirred overnight at
RT. The mixture was added 1N HCl (15 mL) and filtered. The resulted solid was purified
on preparative HPLC to give the product as an off-white solid (280 mg, 65% yield):
HPLC: Waters Symmetry C
18, 5 µm, 3.9 x 150 mm, 1 mL/min, 240 nm, 5% gradient 100% in 5 min, CH
3CN/0.1% H
3PO
4, 4.46 min (97%); mp: 90-92 °C;
1H NMR (DMSO-
d6)
δ 1.01 (t,
J = 7.1 Hz, 6H, CH
3, CH
3), 1.58 (quin,
J = 7.1 Hz, 2H, CH
2), 1.94 - 2.06 (m, 1H, CHH), 2.39 (qd,
J = 4.5, 13.2 Hz, 1H, CHH), 2.55 - 2.73 (m, 7H, CH
2, CH
2, CH
2, CHH), 2.83 - 2.99 (m, 1H, CHH), 3.05 (d,
J = 5.3 Hz, 2H, CH
2), 4.15 - 4.56 (m, 4H, CH
2, CH
2), 5.10 (dd,
J = 5.1, 13.4 Hz, 1H, NCH), 6.17 (br. s., 1H, NH), 6.60 (s, 1H, NH), 7.39 (d,
J = 7.7 Hz, 1H, Ar), 7.45 (s, 1H, Ar), 7.67 (d,
J = 7.9 Hz, 1H, Ar), 8.25 (br. s., 1H, HCOOH), 10.98 (br. s., 1H, NH);
13C NMR (DMSO-
d6)
δ 10.25, 22.49, 26.14, 31.20, 37.39, 42.93, 46.11, 47.09, 51.56, 121.73, 122.81, 126.76,
130.13, 142.27, 145.54, 158.18, 167.99, 171.01, 172.86; LCMS MH
+ = 430; Anal. Calcd. for C
23H
33N
5O
6 +2 H
2O: C, 54.00; H, 7.29; N, 13.69; Found: C, 54.04; H, 6.92; N, 13.63.
5.39 1-[2-(2,6-Dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-3-(4-methyl-tetrahydro-pyran-4-yl)-urea (Example disclosed for reference only)
[0290]

[0291] To the solution of
para-nitrophenyl chloroformate (202 mg, 1 mmol) in CH
3CN (5 mL) was added dropwise at 0 °C a solution of 4-methyl-tetrahydro-pyran-4-ylamine
(150 mg, 1 mmol) and DIPEA (0.082 mL, 1 mmol) in CH
3CN (5 mL). The mixture was stirred at 0 °C for 10 min. 3-(5-Aminomethyl-1-oxo-1,3-dihydro-isoindol-2-yl)-piperidine-2,6-dione
methanesulfonic acid salt (0.37 g, 1 mmol) was added, followed by additional DIPEA
(0.082 mL, 1 mmol). The mixture was allowed to warm to RT and stirred overnight at
RT. The mixture was added 1N HCl (15 mL) and filtered. The resulted solid was purified
on preparative HPLC to give a solid. The solid was recrystalized from DMF (1 mL) and
water (1 mL) to give the product as a white solid (55 mg, 12% yield): HPLC: Waters
Symmetry C
18, 5 µm, 3.9 x 150 mm, 1 mL/min, 240 nm, 5% gradient 100% in 5 min, CH
3CN/0.1% H
3PO
4, 5.02 min (98%); mp: 158-160 °C;
1H NMR (DMSO-
d6)
δ 1.29 (s, 3H, CH
3), 1.40 - 1.58 (m, 2H, CHH), 1.84 - 1.96 (m, 2H, CHH, CHH), 1.99 (s, 1H, CHH), 2.30
- 2.45 (m, 1H, CHH), 2.55 - 2.65 (m, 1H, CHH), 2.84 - 2.99 (m, 1H, CHH), 3.44 - 3.68
(m, 4H, CH
2, CH
2), 4.22 - 4.58 (m, 4H, CH
2, CH
2), 4.95 - 5.29 (m, 1H, NCH), 5.85 (s, 1H, NH), 6.08 - 6.52 (m, 1H, NH), 7.33 - 7.42
(m, 1H, Ar), 7.45 (s, 1H, Ar), 7.68 (d,
J = 7.6 Hz, 1H, Ar), 10.98 (s, 1H, NH);
13C NMR (DMSO-
d6)
δ 22.48, 27.00, 31.20, 36.93, 42.58, 47.10, 48.85, 51.56, 62.92, 120.99, 121.67, 122.88,
126.69, 142.35, 145.48, 157.22, 167.98, 171.01, 172.86; LCMS MH
+ = 415; Anal. Calcd. for C
21H
26N
4O
5 +0.5 H
2O: C, 59.56; H, 6.43; N, 13.23; Found: C, 59.34; H, 6.35; N, 13.21.
5.40 1-(1-Benzyl-piperidin-4-yl)-3-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-urea (Example disclosed for reference only)
[0292]

[0293] To the solution of
para-nitrophenyl chloroformatc (202 mg, 1 mmol) in CH
3CN (5 mL) was added dropwise at 0 °C a solution of 4-amino-benzyl-piperidine (190
mg, 1 mmol) and DIPEA (0.082 mL, 1 mmol) in CH
3CN (5 mL). The mixture was stirred at 0 °C for 10 min. 3-(5-Aminomethyl-1-oxo-1,3-dihydro-isoindol-2-yl)-piperidine-2,6-dione
methanesulfonic acid salt (0.37 g, 1 mmol) was added, followed by additional DIPEA
(0.082 mL, 1 mmol). The mixture was allowed to warm to RT and stirred overnight at
RT. The mixture was added 1N HCl (15 mL) and filtered. The resulted solid was purified
on preparative-HPLC to give a gluey solid. The solid was dissolved in DMF (1 mL) and
saturated NaHCO
3 solution was added dropwise until precipitation started occurring. It was sat still
at RT for 30 min and solid was filtered to give the product as a white solid (55 mg,
12% yield): HPLC: Waters Symmetry C
18, 5 µm, 3.9 x 150 mm, 1 mL/min, 240 nm, 5% gradient 100% in 5 min, CH
3CN/0.1% H
3PO
4, 4.64 min (99%); mp: 208-210 °C;
1H NMR (DMSO-
d6)
δ 1.21 - 1.52 (m, 2H, CHH), 1.63 - 1.82 (m, 2H, CHH), 1.89 - 2.14 (m, 3H, CHH, CHH,
CHH), 2.39 (d,
J = 13.6 Hz, 1H, CHH), 2.55 - 2.65 (m, 1H, CHH), 2.64 - 2.75 (m, 2H, CHH, CHH), 2.83
- 3.02 (m, 1H, CHH), 3.37 - 3.41 (m, 1H, CH), 3.43 (s, 2H, CH
2), 4.21 - 4.50 (m, 3H, CH
2, CH
2), 5.10 (dd,
J = 5.0, 13.3 Hz, 1H, NCH), 5.94 (d,
J = 7.9 Hz, 1H, NH), 6.34 (t,
J = 6.0 Hz, 1H, NH), 7.13 - 7.48 (m, 6H, Ar), 7.66 (d,
J = 7.6 Hz, 1H, Ar), 10.97 (s, 1H, NH);
13C NMR (DMSO-
d6)
δ 22.49, 31.20, 32.46, 42.86, 46.43, 47.09, 51.55, 51.90, 62.19, 121.78, 122.84, 126.79,
128.10, 128.69, 130.13, 138.64, 142.29, 145.44, 157.32, 167.96, 170.99, 172.85; LCMS
MH
+ = 490; Anal. Calcd. for C
21H
26N
4O
5 +0.5 H
2O: C, 59.56; H, 6.43; N, 13.23; Found: C, 59.34; H, 6.35; N, 13.21.
5.41 1-[2-(2,6-Dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-3-piperidin-4-yl-urea hydrochloride salt (Example disclosed for reference only)
[0294]

[0295] Step 1. A mixture of 3-(5-aminomethyl-1-oxo-1,3-dihydro-isoindol-2-yl)-piperidine-2,6-dione
methanesulfonic acid salt (1.25 g, 3.39 mmol) and CDI (0.61 g, 3.73 mmol) in DMF (10
mL) was stirred at RT overnight. 4-Amino-piperidine-1-carboxylic acid
tert-butyl ester (0.68 g, 3.39 mmol) was then added and the mixture was stirred overnight.
The mixture was purified by preparative HPLC to give 4-{3-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1
H-isoindol-5-ylmethyl]-ureido}-piperidine-1-carboxylic acid
tert-butyl ester as an off-white solid (0.46 g, 27% yield). The product was used in the
next step without further purification. HPLC, Waters Symmetry C
18, 5 µm, 3.9 x 150 mm, 1 ml/min, 240 nm, 30/70 CH
3CN/0.1% H
3PO
4, 4.35 min (99.4%).
[0296] Step 2: A mixture of 4-{3-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-ureido}-piperidine-1-carboxylic
acid
tert-butyl ester (0.45 g, 0.90 mmol) in 2 M hydrochloride in diethyl ether (30 mL, 9.01
mmol) was stirred overnight. The white suspension was filtered and washed with diethyl
ether. The resulting white solid was stirred in acetonitrile (80 mL) at 50 °C overnight.
The suspension was filtered, washed with acetonitrile, and vacuum dried to give the
product as a white solid (0.34 g, 86% yield): HPLC:Waters Xterra C
18, 5 µm, 3.9 x 150 mm, 1 ml/min, 240 nm, 5/95 CH
3CN/0.1% HCOONH
4 gradient to 95/5 in 5 min, kept 5 min, 3.10 min (95.1%); mp, 340-342 °C;
1H NMR (DMSO-
d6)
δ 1.46 - 1.65 (m, 2H, CH
H, CH
H), 1.85 - 2.05 (m, 3H, CH
H, CH
H, CH
H), 2.30 - 2.47 (m, 1H, CH
H), 2.55 - 2.67 (m, 1H, CH
H), 2.83 - 3.03 (m, 3H, CH
H, CH
H, CH
H), 3.15 - 3.27 (m, 2H, CH
H, CH
H), 3.59 - 3.77 (m, 1H, C
H), 4.23 - 4.51 (m, 4H, C
H2, C
H2), 5.11 (dd,
J = 4.9, 13.2 Hz, 1H, NC
H), 6.40 - 6.57 (m, 2H, N
H, N
H), 7.39 (d,
J = 7.9 Hz, 1H, Ar
H), 7.45 (s, 1H, Ar
H), 7.67 (d,
J = 7.7 Hz, 1H, Ar
H), 8.75 (br. s., 2H, Cl
H2N), 10.98 (s, 1H, N
H);
13C NMR (DMSO-
d6)
δ 22.49, 28.92, 31.20, 41.96, 42.82, 43.95, 47.09, 51.55, 121.76, 122.84, 126.76, 130.14,
142.29, 145.35, 157.32, 167.98, 171.01, 172.87; LC/MS MH
+ = 400; Anal. Calcd. For C
20H
26N
5O
4Cl +1.7 H
2O: C, 51.49; H, 6.35; N, 15.01; Cl, 7.60; Found: C, 51.18; H, 6.15; N, 14.90; Cl,
7.51.
5.42 Isoindolin compounds (Example disclosed for reference only)
[0297]

[0298] The isoindolin compounds shown above are using the procedure as described in Section
5.15.
5.43 4-{3-[2-(2,6-Dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-ureido}-cyclohexanecarboxylic
acid amide (Example disclosed for reference only)
[0299]

[0300] To the solution of para-nitro-phenyl chloroformate (200 mg, 1 mmol) in CH
3CN (5 mL), was added the CH
3CN solution (5 mL) of 4-Amino-cyclohexanecarboxylic acid amide (210 mg, 1 mmol) and
DIPEA (0.3 mL, 2 mmol) dropwise at room temperature. The mixture was stirred at room
temperature for 10 min. 3-(5-Aminomethyl-1-oxo-1,3-dihydro-isoindol-2-yl)-piperidine-2,6-dione
methanesulfonic acid salt (0.37 g, 1 mmol) was added followed by additional DIPEA
(0.15 mL, 1 mmol). The mixture was stirred overnight at ambient temperature. The mixture
was then filtered. The filtrate was purified on prep-HPLC to give 4-{3-[2-(2,6-dioxo-piperidin-3-yl)-1-oxy-2,3-dihydro-1H-isoindol-5-ylmethyl]-ureido}-cyclohexanecarboxylic
acid amide as a white solid (130 mg, 30% yield). HPLC: Waters Symmetry C
18, 5µm, 3.9 x 150 mm, 1 mL/min, 240 nm, 5% grad 95% in 5 min, CH
3CN/0.1% H
3PO
4,: t
R =4.78 min (92%); mp: 195-197 °C;
1H NMR (DMSO-d
6) δ 1.32 - 1.73 (m, 8H, CHH+CHH+CHH+CHH), 1.97 (br. S., 1H, CHH), 2.09 - 2.21 (m,
1H, CHH), 2.40 (d,
J = 13.4 Hz, 1H, CHH), 2.62 (br. S., 1H, CHH), 2.80 - 3.03 (m, 1H, CHH), 3.70 (br.
S., 1H, CHH), 4.18 - 4.58 (m, 4H, CH
2 + CH
2), 5.11 (dd, 1H, CHN), 6.15 (d,
J = 7.9 Hz, 1H, NH), 6.38 (t,
J = 5.9 Hz, 1H, NH), 6.67 (br. S., 1H, NH), 7.19 (br. S., 1H, NH), 7.38 (d,
J = 7.7 Hz, 1H, Ar), 7.45 (s, 1H, Ar), 7.67 (d,
J = 7.9 Hz, 1H, Ar), 10.98 (br. S., 1H, NH);
13C NMR (DMSO-d
6) δ 0.03, 22.42, 24.06, 24.19, 29.73, 41.73, 42.79, 44.16, 47.03, 51.49, 121.68, 122.80,
126.71, 130.09, 142.26, 145.37, 157.28, 163.37, 167.89, 170.92, 172.78, 176.83; LC-MS:
442; Anal Calcd for C
22H
27N
5O
5 +0.5 HCOOH+ 1 H
2O + 0.2 CH
3CN: C, 56.05; H, 6.21; N, 14.84; Found: C, 55.70; H, 6.46; N, 14.78.
5.44 1-(6-Chloro-pyridin-3-yl)-3-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-urea (Example disclosed for reference only)
[0301]

[0302] To a suspension of 3-(5-aminomethyl-1-oxo-1,3-dihydro-isoindol-2-yl)-piperidine-2,6-dione
methanesulfonic acid salt (0.37 g, 1 mmol) in DMF (3 mL) was added carbonyl diimidazole
(162 mg, 1 mmol). The mixture was stirred at room temperature overnight. 5-Amino-2-chloropyridine
(128.5 mg, 1 mmol) was added to the mixture and the mixture was heated at 80 °C for
6 hours. The mixture was then concentrated under reduced pressure and the resulted
mixture was purified on silica gel column eluted with methylene chloride and methanol
to give 11-(6-chloro-pyridin-3-yl)-3-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-urea
as an off-white solid (20 mg, 5% yield): HPLC: Waters Symmetry C
18, 5µm, 3.9 x 150 mm, 1 mL/min, 240 nm, 5% grad 95% in 5 min, CH
3CN/0.1% H
3PO
4, t
R = 5.43 min (97%); mp: 224-226 °C;
1H NMR (DMSO-d
6) δ 1.89 - 2.09 (m, 1H, CHH), 2.29 - 2.44 (m, 1H, CHH), 2.54 - 2.66 (m, 1H, CHH),
2.83 - 3.04 (m, 1H, CHH), 4.20 - 4.57 (m, 4H, CH
2,CH
2), 5.11 (dd,
J = 5.0, 13.3 Hz, 1H, NCH), 6.99 (t,
J = 6.2 Hz, 1H, NH), 7.37 (d,
J = 8.5 Hz, 1H, Ar), 7.45 (d,
J = 7.7 Hz, 1H, Ar), 7.52 (s, 1H, Ar), 7.70 (d,
J = 7.7 Hz, 1H, Ar), 7.95 (dd,
J = 2.8, 8.7 Hz, 1H, Ar), 8.42 (d,
J = 2.3 Hz, 1H, Ar), 9.03 (s, 1H, NH), 10.98 (s, 1H, NH);
13C NMR (DMSO-d
6) δ 22.49, 31.18, 42.86, 47.12, 51.58, 121.91, 122.94, 123.89, 126.92, 128.37, 130.42,
136.75, 139.04, 142.39, 144.50, 154.97, 167.93, 171.01, 172.86, 215.61; LC-MS: 428;
Anal Calcd for C
20H
18N
5O
4Cl +0.1 H
2O C, 55.91; H, 4.27; N,16.30; Cl, 8.25; Found: C, 56.31; H, 4.51; N, 15.92; Cl, 8.47.
5.45 1-[4-(2,4-Difluoro-phenyl)-thiazol-2-yl]-3-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-urea (Example disclosed for reference only)
[0303]

[0304] A mixture of 3-(5-aminomethyl-1-oxo-1,3-dihydro-isoindol-2-yl)-piperidine-2,6-dione
methanesulfonic acid salt (0.40 g, 1.1 mmol) and CDI (0.18 g, 1.1 mmol) in DMF (5
mL) was stirred at room temperature for 18h. To the mixture was added 4-(2,4-difluoro-phenyl)-thiazol-2-ylamine
(0.23 g, 1.1 mmol) at room temperature, and the mixture was stirred at 100 °C for
2 days. To the mixture was added water (25 mL) and ethyl acetate (20 mL). The mixture
was stirred at room temperature for 2 h. The suspension was filtered and the solid
was washed with water (20 mL), ethyl acetate (20 mL) and water (20 mL) to give a solid.
The solid was purified with Prep HPLC to give 1-[4-(2,4-difluoro-phenyl)-thiazol-2-yl]-3-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-urea
as a white solid (70 mg, 13% yield): HPLC: Waters Symmetry C18, 5µm, 3.9 x 150 mm,
1 mL/min, 240 nm, 40/60 CH
3CN/0.1% H
3PO
4, 6.43 min (97.7 %); mp: 264-266 °C; 1H NMR (DMSO-d6) 1.93 - 2.05 (m, 1H, CHH), 2.30
- 2.45 (m, 1H, CHH), 2.55 - 2.68 (m, 1H, CHH), 2.82 - 3.00 (m, 1H, CHH), 4.32 (d,
J = 17.4 Hz, 1H, CHH), 4.41 - 4.54 (m, 3H, CH2, CHH), 5.11 (dd, J = 5.1, 13.2 Hz,
1H, NCH), 7.09 - 7.24 (m, 2H, Ar, NH), 7.28 - 7.41 (m, 2H, Ar), 7.46 (d, J = 7.7 Hz,
1H, Ar), 7.53 (s, 1H, Ar), 7.71 (d, J = 7.9 Hz, 1H, Ar), 7.93 - 8.10 (m, 1H, Ar),
10.85 (s, 1H, NH), 10.98 (s, 1H, NH); 13C NMR (DMSO-d6) 22.48, 31.18, 42.94, 47.12,
51.57, 104.54 (t, JC-F = 26.4 Hz), 110.67 (d, JC-F = 14.3 Hz), 111.81 (dd, JC-F =
3.3, 20.9 Hz), 118.91 (dd, JC-F = 2.8, 11.6 Hz), 121.95, 123.03, 126.95, 130.29 (dd,
JC-F = 4.4, 9.9 Hz), 130.49, 141.44, 142.45, 143.95, 154.02, 159.48 (dd, JC-F = 252.2,
12.1 Hz), 159.36, 161.24 (dd, JC-F = 246.9, 13.2 Hz), 167.88, 170.98, 172.85; LCMS
MH = 512; Anal. Calcd for C24H19N5O4F2S: C, 56.36; H, 3.74; N, 13.69; Found: C, 56.16;
H, 3.80; N, 14.07.
5.46 6-{3-[2-(2,6-Dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-ureido}-3,4-dihydro-1H-isoquinoline-2-carboxylic
acid tert-butyl ester (Example disclosed for reference only)
[0305]

[0306] 3-(5-aminomethyl-1-oxo-1,3-dihydro-isoindol-2-yl)-piperidine-2,6-dione methanesulfonate
(1.11 g, 3.0 mmol) and 1,1'-Carbonyldiimidazole (535 mg, 3.3 mmol) were suspended
in dry DMF (20 mL) and the mixture was stirred at rt for 24 h. While stirring, a portion
of the reaction mixture (6.7 mL, ∼1 mmol) was transferred to a vial containing tert-butyl
6-amino-3,4-dihydroisoquinoline-2(1H)-carboxylate (273 mg, 1.1 mmol). The resulting
mixture was stirred at rt overnight and the reaction progress was monitored by LCMS.
After 48 h, additional tert-butyl 6-amino-3,4-dihydroisoquinoline-2(1H)-carboxylate
(50 mg, 0.2 mmol) was transferred to the reaction mixture and stirring continued for
another 24 h. The reaction mixture was acidified with acidic acid and water. The volatiles
were removed in vacuo and the residue was dissolved in DMF and purified using C-18
preparatory HPLC to give 6-{3-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-ureido}-3,4-dihydro-1H-isoquinoline-2-carboxylic
acid tert-butyl ester as a white solid (290 mg, 53% yield): HPLC:Waters Symmetry C
18, 5 µm, 3.9 x 150 mm, 1 ml/min, 240 nm, 40/60 CH
3CN/0.1% H
3PO
4, 5.02 min (96.5%); mp: 230-232 °C;
1H NMR (DMSO-d
6) δ 1.42 (s, 9H, (CH
3)
3), 1.90 - 2.11 (m, 1H, CHH), 2.23 - 2.47 (m, 1H, CHH), 2.60 (d,
J = 17.9 Hz, 1H, CHH), 2.70 (t,
J = 5.6 Hz, 2H, CH
2), 2.81 - 3.06 (m, 1H, CHH), 3.51 (t,
J = 5.7 Hz, 2H, CH
2), 4.18 - 4.58 (m, 6H, CH
2, CH
2, CH
2), 5.11 (dd,
J = 4.9, 13.2 Hz, 1H, CH), 6.72 (t,
J = 5.9 Hz, 1H, NH), 7.00 (d,
J = 8.3 Hz, 1H, Ar), 7.17 (d,
J = 8.1 Hz, 1H, Ar), 7.27 (br. s., 1H, Ar), 7.44 (d,
J = 7.9 Hz, 1H, Ar), 7.51 (s, 1H, Ar), 7.69 (d,
J = 7.9 Hz, 1H, Ar), 8.56 (s, 1H, NH), 10.98 (s, 1H, NH);
13C NMR (DMSO-d
6) δ 22.46, 28.07, 28.44, 31.16, 41.51, 42.75, 45.04, 47.08, 51.55, 78.81, 116.01,
117.49, 121.87, 122.90, 126.20, 126.36, 126.88, 130.25, 134.70, 138.56, 142.35, 144.86,
153.97, 155.21, 167.93, 170.96, 172.83; LCMS: MH = 548; Anal Calcd for C
29H
33N
5O
6 + 1.1 H
2O: C, 61.39; H, 6.25; N, 12.34. Found: C, 61.38; H, 6.11; N, 12.29.
5.47 Isoindolin compounds of Formula III (Example disclosed for reference only)
[0307]

[0308] The isoindolin compounds of Formula III are made as shown in Scheme 1.

[0309] To a solution of the 5-aminomethylisoindolinone glutarimide 1 (1.134 mmol) in dry
DMF (10 mL) is added Cs
2CO
3 (406 mg, 1.248 mmol). After 5 minutes of stirring, an appropriate chloromethoxycarbonyl
reagent 2 (1.248 mmol) is added and the resulting mixture is stirred at 50 °C overnight.
The reaction mixture is neutralized with 0.5N HCl and then concentrated to dryness
in vacuo. The residue is purified by preparative HPLC, followed by the removal of protecting
groups, if necessary, under standard conditions to yield a compound of Formula III.
5.48 2,2-Dimethyl-propionic acid 3-{5-[3-(3-chloro-4-methyl-phenyl)-ureidomethyl]-1-oxo-1,3-dihydro-isoindol-2-yl}-2,6-dioxo-piperidin-1-ylmethyl
ester (Example disclosed for reference only)
[0310]

[0311] To a solution of 1-(3-chloro-4-methyl-phenyl)-3-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1
H-isoindol-5-ylmethyl]-urea (500 mg, 1.134 mmol) in dry DMF (10 mL) was added Cs
2CO
3 (406 mg, 1.248 mmol). After 5 min of stirring, chloromethyl pivalate (188 mg, 1.248
mmol) was added and the resulting mixture was stirred at 50 °C overnight. The reaction
mixture was neutralized with 0.5N HCl and then concentrated to dryness
in vacuo. The residue was dissolved in acetonitrile and water was added slowly resulting in
a precipitate to form. The solid was collected by filtration and dissolved in DMF
for purification on a C-18 preparative HPLC column (50/50 CH
3CN/water). Fractions containing the desired product were combined, concentrated
in vacuo to remove most of the CH
3CN, and then lyophilized to provide the product as a white solid (81 mg, 13% yield):HPLC:
Waters Symmetry C
18, 5 µm, 3.9 x 150 mm, 1 mL/min, 240 nm, 55/45 CH
3CN/0.1% H
3PO
4, 4.58 min (97.2 %); mp: 155-157 °C;
1H NMR (DMSO-d
6)
δ 0.95 - 1.23 (m, 9H,
tBu), 1.96 - 2.15 (m, 1H, CH), 2.23 (s, 3H, CH
3), 2.31 - 2.47 (m, 1H, CH), 2.83 (d,
J = 16.6 Hz, 1H, CH), 3.00 - 3.22 (m, 1H, CH), 4.17 - 4.60 (m, 4H, CH
2, CH
2), 5.30 (dd,
J = 5.0, 13.3 Hz, 1H, CH), 5.47 - 5.83 (m, 2H, CH
2), 6.79 (t,
J = 5.9 Hz, 1H, CH
2NH), 7.02 - 7.28 (m, 2H, Ar), 7.36 - 7.60 (m, 2H, Ar), 7.60 - 7.86 (m, 2H, Ar), 8.74
(s, 1H, NH);
13C NMR (DMSO-
d6)
δ 18.73, 21.56, 26.62, 31.21, 38.21, 42.79, 47.04, 52.01, 63.39, 116.45, 117.66, 121.95,
123.01, 126.98, 127.42, 130.11, 131.00, 132.99, 139.58, 142.36, 144.87, 155.07, 167.96,
170.13, 171.23, 176.51; LCMS: MH
+ = 555, 557; Anal. Calcd. for C
28H
31ClN
4O
6 + 0.2 H
2O: C, 60.20; H, 5.67; N, 10.03; Cl, 6.35; Found: C, 60.20; H, 5.74; N, 9.94; Cl, 6.39.
5.49 (3-(5-((3-(3-Chloro-4-methylphenyl)ureido)methyl)-1-oxoisoindolin-2-yl)-2,6-dioxopiperidin-1-ylmethyl
piperidine-4-carboxylate hydrochloride (Example disclosed for reference only)
[0312]

[0313] To 1-tert-butyl 4-(3-(5-((3-(3-chloro-4-methylphenyl)ureido)methyl)-1-oxoisoindolin-2-yl)-2,6-dioxopiperidin-1-yl)methyl
piperidine-1,4-dicarboxylate (300 mg, 0.44 mmol, obtained as described in previous
example) was added 2 N HCl in Et
2O (15 mL). The slurry was vigorously stirred for 2 h at rt. The solid was filtered,
washed with copious Et
2O, and dried in a vacuum oven overnight to give (3-(5-((3-(3-chloro-4-methylphenyl)ureido)methyl)-1-oxoisoindolin-2-yl)-2,6-dioxopiperidin-1-yl)methyl
piperidine-4-carboxylate Hydrochloride as a white solid (285 mg, 105% yield): HPLC:Waters
Symmetry C
18, 5 µm, 3.9 x 150 mm, 1 ml/min, 240 nm, 30/70 CH
3CN/0.1% H
3PO
4, 3.26 min (98.2%); mp: 234-236 °C;
1H NMR (DMSO-d
6) δ 1.62 - 1.81 (m, 2H, CHH, CHH), 1.87 - 2.01 (m, 2H, CHH, CHH), 2.02 - 2.16 (m,
1H, CHH), 2.16 - 2.29 (m, 3H, CH
3), 2.32 - 2.47 (m, 1H, CHH), 2.59 - 2.76 (m, 1H, CHH), 2.77 - 2.99 (m, 3H, CHH, CHH,
CHH), 3.00 - 3.28 (m, 3H, CHH, CHH, CHH), 4.15 - 4.66 (m, 4H, CH
2, CH
2), 5.26 (dd,
J = 4.9, 13.2 Hz, 1H, CH), 5.55 - 5.80 (m, 2H, CH
2O), 7.02 (t,
J = 5.9 Hz, 1H, NH), 7.09 - 7.29 (m, 2H, Ar), 7.45 (d,
J = 7.9 Hz, 1H, Ar), 7.53 (s, 1H, Ar), 7.62 - 7.82 (m, 2H, Ar), 8.41 - 8.96 (m, 2H,
NH, NH), 9.09 (s, 1H, NH);
13C NMR (DMSO-d
6) δ 18.73, 21.46, 24.31, 31.24, 37.30, 42.00, 42.73, 47.20, 52.09, 63.40, 116.31,
117.50, 121.91, 122.99, 126.97, 127.27, 130.09, 130.99, 132.96, 139.69, 142.38, 144.94,
155.21, 167.96, 170.19, 171.29, 172.20; LCMS: MH = 582, 584; Anal Calcd for C
29H
33Cl
2N
5O
6 + 1.6H
2O + 0.6 HCl + 0.2 Et
2O + 0.4
tBuCl: C, 53.35; H, 6.05; N, 9.91; Cl, 13.04. Found: C, 53.13; H, 6.06; N, 9.70; Cl,
13.25.
5.50 1-tert-Butyl 4-(3-(5-((3-(3-chloro-4-methylphenyl)ureido)methyl)-1-oxoisoindolin-2-yl)-2,6-dioxopiperidin-1-yl)methyl
piperidine-1,4-dicarboxylate (Example disclosed for reference only)
[0314]

Step 1: 1-tert-Butyl 4-chloromethyl piperidine-1,4-dicarboxylate
[0315]

[0316] To a mixture of 1-(tert-butoxycarbonyl)piperidine-4-carboxylic acid (3 g, 13.1 mmol)
in dichloromethane (40 mL) and water (50 mL)was added NaHCO
3 (4.4 g, 52.3 mmol) and tetrabutylammonium hydrogen sulfate (444 mg, 1.3 mmol). After
stirring the mixture in an ice bath at 0 °C for ∼10 min, chloromethyl chlorosulfate
(2.59 g, 15.7 mmol) in 10 mL of dichloromethane was added dropwise. The reaction mixture
was allowed to warm up to rt and stirred vigorously overnight. The mixture was transferred
to a separatory funnel with dichloromethane and water (200 mL, each). The organic
layer was washed with additional water and brine, dried (Na
2SO
4), and concentrated in vacuo to give 1-tert-butyl 4-chloromethyl piperidine-1,4-dicarboxylate
as a clear oil (3.5 g, 96% yield). This material was used without further purification.
1H NMR (DMSO-d
6) δ 1.26 - 1.48 (m, 11H, CHH, CHH, tertBu), 1.83 (dd,
J = 3.0, 13.2 Hz, 2H, CHH, CHH), 2.58 - 2.75 (m, 1H, CH), 2.74 - 2.99 (m, 2H, CHH,
CHH), 3.61 - 4.02 (m, 2H, CHH, CHH), 5.87 (s, 2H, CH
2O);
13C NMR (DMSO-d
6) δ 27.16, 27.98, 39.58, 42.35, 69.41, 78.67, 153.75, 172.25; LCMS: MH = 278.
Step 2: 1-tert-Butyl 4-iodomethyl piperidine-1,4-dicarboxylate
[0317]

[0318] To a solution of NaI (1.6 g, 11.09 mmol) in dry acetonitrile (8 mL) was added dropwise
1-tert-butyl 4-chloromethyl piperidine-1,4-dicarboxylate (2.8 g, 10.08 mmol) in 2
mL of acetonitrile. The mixture turned dark orange and was stirred at rt in the dark
for 24 h. The reaction mixture was filtered to remove NaCl and the filtrate was concentrated
in vacuo. The remaining residue was partitioned between DCM and 5% aq NaHSO
3 solution. The organic layer was washed with water and brine, dried (Na
2SO
4), and concentrated in vacuo to give 1-tert-butyl 4-iodomethyl piperidine-1,4-dicarboxylate
as a pale yellow oil (3.5 g, 94%). Attempts to characterize by NMR in DMSO-d
6 resulted in rapid decomposition. The material was stored in the dark at -20 °C to
minimize decomposition before further use. LCMS: MH = 370.
Step 3: 1-tert-Butyl 4-(3-(5-((3-(3-chloro-4-methylphenyl)ureido)methyl)-1-oxoisoindolin-2-yl)-2,6-dioxopiperidin-1-yl)methyl
piperidine-1,4-dicarboxylate
[0319]

[0320] 1-(3-Chloro-4-methyl-phenyl)-3-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-urea
(400 mg, 0.907 mmol) and NaH (43.5 mg, 1.82 mmol) were stirred in dry DMF (15 mL)
for ∼ 10 min. To the mixture was added 1-tert-butyl 4-iodomethyl piperidine-1,4-dicarboxylate
(402 mg, 1.09 mmol) and the reaction stirred in the dark at rt for 2 h. The reaction
was quenched with acetic acid (5 mL) and concentrated in vacuo. The residue was partitioned
in water and EtOAc (100 mL, each). The separated organic layer was washed with additional
water and brine, dried (Na
2SO
4), and concentrated in vacuo to give 1.4 gm of a yellowish solid which was dissolved
in DMF for purification on a C-18 prep HPLC column. Fractions containing the desired
product were combined, concentrated
in vacuo to remove most of the CH
3CN, and then lyophilized to provide 1-tert-butyl 4-(3-(5-((3-(3-chloro-4-methylphenyl)ureido)methyl)-1-oxoisoindolin-2-yl)-2,6-dioxopiperidin-1-yl)methyl
piperidine-1,4-dicarboxylate as a white solid (380 mg, 49% of theoretical yield from
two separate runs that were combined before prep HPLC). HPLC:Waters Symmetry C
18, 5 µm, 3.9 x 150 mm, 1 ml/min, 240 nm, 55/45 CH
3CN/0.1% H
3PO
4, 5.82 min (99.4%); mp: 157-159 °C;
1H NMR (DMSO-d
6) δ 1.23 - 1.51 (m, 11H, (CH
3)
3, CHH, CHH), 1.68 - 1.84 (m, 2H, CHH, CHH), 1.97 - 2.13 (m, 1H, CHH), 2.15 - 2.31
(m, 4H, CH
3, CH), 2.32 - 2.46 (m, 1H, CHH), 2.68 - 2.94 (m, 3H, CHH, CHH, CHH), 2.98 - 3.21 (m,
1H, CHH), 3.69 - 3.89 (m, 2H, CHH, CHH), 4.20 - 4.57 (m, 4H, CH
2, CH
2), 5.27 (dd,
J = 5.0, 13.3 Hz, 1H, CH), 5.53 - 5.73 (m, 2H, CH
2O), 6.81 (t,
J = 5.9 Hz, 1H, NH), 7.06 - 7.24 (m, 2H, Ar), 7.45 (d,
J = 7.9 Hz, 1H, Ar), 7.53 (s, 1H, NH), 7.67 (d,
J = 1.9 Hz, 1H, Ar), 7.71 (d,
J = 7.7 Hz, 1H, Ar), 8.76 (s, 1H, NH);
13C NMR (DMSO-d
6) δ 18.66, 21.39, 27.38, 27.97, 31.16, 42.72, 47.08, 51.98, 63.15, 78.60, 116.37,
117.60, 121.88, 122.93, 126.91, 127.34, 130.06, 130.94, 132.91, 139.52, 142.32, 144.82,
153.74, 155.01, 167.90, 170.11, 171.18, 172.97 (two
13C signals arising from the piperidine ring are not observed due to overlap with DMSO-d
6); LCMS: MH = 682, 684; Anal Calcd for C
34H
40ClN
5O
8 + 1.0 H
2O: C, 58.32; H, 6.05; N, 10.00; Cl, 5.06. Found: C, 58.41; H, 5.93; N, 9.96; Cl, 5.22.
5.51 N-[2-(2,6-Dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-4-imidazol-1-yl-benzamide (Example disclosed for reference only)
[0321]

[0322] A mixture of 4-imidazol-1-yl-benzoic acid (0.21 g, 1.1 mmol) and CDI (0.19 g, 1.2
mmol) in DMF (14 mL) was stirred at 40 °C for 2 hrs. To the suspension was added 3-(5-aminomethyl-1-oxo-1,3-dihydro-isoindol-2-yl)-piperidine-2,6-dione
methanesulfonic acid salt (0.40 g, 1.1 mmol) and the mixture was stirred at 40 °C
for 20 hrs. Water (20 mL) was added to the reaction mixture and the mixture was stirred
at RT for 30 min. The suspension was filtered and the solid was washed with water
(20 mL), ethyl acetate (20 mL), and water (20 mL) to give the product as a white solid
(340 mg, 71% yield): HPLC: Waters Xterra C
18, 5 µm, 3.9 x 150 mm, 1 mL/min, 240 nm, 5% gradient 95% in 5 min, CH
3CN/0.1% NH
4OOCH, 3.92 min (99.0 %); mp: 268-270 °C;
1H NMR (DMSO-
d6)
δ 1.90 - 2.08 (m, 1H, CHH), 2.24 - 2.47 (m, 1H, CHH), 2.54 - 2.67 (m, 1H, CHH), 2.82
- 3.02 (m, 1H, CHH), 4.31
(d, J= 17.4 Hz, 1H, CHH), 4.45 (d,
J = 17.4 Hz, 1H, CHH), 4.62 (d,
J = 5.7 Hz, 2H, CH
2), 5.11 (dd,
J = 5.1, 13.2 Hz, 1H,NCH), 7.14 (s, 1H, Ar), 7.49 (d,
J = 7.9 Hz, 1H, Ar), 7.56 (s, 1H, Ar), 7.71 (d,
J = 7.7 Hz, 1H, Ar), 7.77 - 7.91 (m, 3H, Ar), 8.05 (d,
J = 8.7 Hz, 2H, Ar), 8.39 (s, 1H, Ar), 9.24 (t,
J = 5.9 Hz, 1H, NH), 10.98 (br. s., 1H, NH);
13C NMR (DMSO-
d6)
δ 22.49, 31.20, 42.79, 47.13, 51.58, 117.78, 119.59, 122.10, 122.97, 127.08, 129.02,
130.20, 130.39, 132.07, 135.60, 138.99, 142.40, 143.83, 165.25, 167.92, 170.99, 172.86;
LCMS: MH
+ = 444; Anal. Calcd. for C
24H
21N
5O
4 + 1.5 H
2O: C, 61.27; H, 5.14; N, 14.89; Found: C, 61.03; H, 4.93; N, 14.75.
5.52 N-[2-(2,6-Dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-4-morpholin-4-yl-benzamide (Example disclosed for reference only)
[0323]

[0324] A mixture of 4-morpholin-4-yl-bcnzoic acid (0.22 g, 1.1 mmol) and CDI (0.19 g, 1.2
mmol) in DMF (4 mL) was stirred at 40 °C for 2 hrs. To the suspension was added 3-(5-aminomethyl-1-oxo-1,3-dihydro-isoindol-2-yl)-piperidine-2,6-dione
methanesulfonic acid salt (0.40 g, 1.1 mmol), and the mixture was stirred at 40 °C
for 2 hrs. Water (20 mL) was added to the reaction mixture and the mixture was stirred
at RT for 30 min. The suspension was filtered and the solid was washed with water
(20 mL), ethyl acetate (20 mL), and water (20 mL) to give the product as a white solid
(370 mg, 74% yield): HPLC: Waters Xterra C
18, 5 µm, 3.9 x 150 mm, 1 mL/min, 240 nm, 5% gradient 95% in 5 min, CH
3CN/0.1% NH4OOCH, 1.10 min (96.5 %); mp: 275-277 °C;
1H NMR (DMSO-
d6) δ 2.00 (d,
J = 5.1 Hz, 1H, CHH), 2.38 (m, 1H, CHH), 2.54 - 2.66 (m, 1H, CHH), 2.81 - 3.01 (m,
1H, CHH), 3.15 - 3.28 (m, 4H, CH
2, CH
2), 3.65 - 3.80 (m, 4H, CH
2, CH
2), 4.30 (d,
J = 17.4 Hz, 1H, CHH), 4.44 (d,
J = 17.6 Hz, 1H, CHH), 4.56 (d,
J = 5.9 Hz, 2H, CH
2), 5.10 (dd,
J = 5.1, 13.2 Hz, 1H, NCH), 6.98 (d,
J = 9.1 Hz, 2H, Ar), 7.45 (d,
J = 7.9 Hz, 1H, Ar), 7.51 (s, 1H, Ar), 7.68 (d,
J = 7.9 Hz, 1H, Ar), 7.81
(d, J= 8.9 Hz, 2H, Ar), 8.88 (s, 1H, NH), 10.98 (s, 1H, NH);
13C NMR (DMSO-
d6)
δ 22.46, 31.15, 42.54, 47.07, 47.33, 51.53, 65.86, 113.40, 121.96, 122.86, 123.72,
126.99, 128.50, 130.24, 142.31, 144.38, 152.95, 165.87, 167.91, 170.96, 172.83; LCMS:
MH
+ = 463; Anal. Calcd. for C
25H
26N
4O
5 + 0.6 H
2O: C, 63.44; H, 5.79; N, 11.84; Found: C, 63.19; H, 5.42; N, 12.09.
5.53 2-Amino-2-(4-chloropheyl)-N-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)acetamide hydrochloride (Example disclosed for reference only)
[0325]

[0326] Step 1: To a stirred solution of
N-Boc-(4'-chlorophenyl) glycine (1.16 g, 4.10 mmol) in DMF (20 mL) was added CDI (0.69
g, 4.30 mmol). The reaction mixture was heated to 40°C for 2 hrs. 3-(5-Aminomethyl-1-oxo-1,3-dihydro-isoindol-2-yl)-piperidine-2,6-dione
hydrochloride (1.50 g, 4.10 mmol) was then added. After 3.5 hrs at 40 °C, water (70
mL) was added. Solid precipitated was isolated by filtration, washed with water (3
x 20 mL), and
dried in vacuo to afford
tert-butyl 1-(4-chlorophenyl)-2-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methylamino)-2-oxoethylcarbamate
as a white solid (1.77 g, 80% yield). The crude product was used in the next step
without further purification.
[0327] Step 2: To a stirred solution of
tert-butyl 1-(4-chlorophenyl)-2-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methylamino)-2-oxoethylcarbamate
(0.84 g, 1.50 mmol) in EtOAc (35 mL) was added a solution of diluted HCl (2N in ether,
30 mL). After 3 days, solid was isolated by filtration, washed with ether (3 X 10
mL), and dried
in vacuo. The solid was then dissolved in water (100 mL) and extracted with EtOAc (2 x 50 mL).
The aqueous phase was concentrated and the residue was triturated with ether for 1
hr. The product was isolated by filtration and dried
in vacuo to give the product as a white solid (0.65 g, 88% yield): HPLC: Waters Symmetry C
18, 5 µm, 3.9 x 150 mm, 1 mL/min, 240 nm, gradient: CH
3CN/0.1% H
3PO
4: 10/90 to 90/10 in 10 min, 90/10 (5 min): 4.91 min (98.00%); mp: 255-257 °C;
1H NMR (DMSO-
d6)
δ 1.93 - 2.09 (m, 1H, CHH), 2.39 (qd,
J = 4.4, 13.1 Hz, 1H, CHH), 2.54 - 2.69 (m, 1H, CHH), 2.82 - 3.03 (m, 1H, CHH), 4.24
(d,
J = 17.4 Hz, 1H, CHH), 4.30 - 4.54 (m, 3H, CH
2, CHH), 5.02 - 5.19 (m, 2H, CH, CH), 7.22 - 7.41 (m, 2H, Ar), 7.47 - 7.71 (m, 5H,
Ar), 9.35 (t,
J = 5.9 Hz, 1H, NH), 10.99 (s, 1H, NH);
13C NMR (DMSO-
d6)
δ 22.48, 31.20, 42.26, 47.03, 51.59, 54.80, 121.89, 121.96, 122.85, 126.97, 128.80,
129.81, 130.49, 133.02, 133.99, 142.27, 142.70, 167.22, 167.82, 170.98, 172.85; LCMS:
MH
+ = 441/443; Anal. Calcd. for C
22H
22N
4O
4Cl
2 + 0.6 H
2O: C, 54.13; H, 4.79; N, 11.48; Found: C, 53.89; H, 4.82; N, 11.60.
5.54 2-Amino-5-chloro-N-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-benzamide (Example disclosed for reference only)
[0328]

[0329] DIPEA (1.46 mL, 8.12 mmol) was added to a stirred mixture of 3-(5-aminomethyl-1-oxo-1,3-dihydro-isoindol-2-yl)-piperidine-2,6-dione
methanesulfonate (1.0 g, 2.71 mmol) and 5-chloro-isatoic anhydride 3 (0.535 g, 2.71
mmol) in acetonitrile (10 mL) under nitrogen. The reaction mixture was refluxed for
14 hrs, at which time LCMS indicated the reaction was complete. Water was added to
the slurry and the solid was collected by filtration, washed with additional water,
and dried
in vacuo to provide the product as a whte solid (860 mg, 74% yield): HPLC: Waters Symmetry
C
18, 5 µm, 3.9 x 150 mm, 1 mL/min, 240 nm, 30/70 CH
3CN/0.1% H
3PO
4, 6.30 min (98.7 %); mp: 242-244 °C;
1H NMR (DMSO-
d6)
δ 1.79 - 2.16 (m, 1H, CHH), 2.29 - 2.46 (m, 1H, CHH), 2.60 (d,
J = 16.6 Hz, 1H, CHH), 2.74 - 3.08 (m, 1H, CHH), 4.08 - 4.69 (m, 4H, CH
2, CH
2), 5.11 (dd,
J = 4.9, 13.2 Hz, 1H, CH), 6.60 (s, 2H, NH
2), 6.73 (d,
J = 8.7 Hz, 1H, Ar), 7.18 (dd,
J = 2.4, 8.8 Hz, 1H, Ar), 7.46 (d,
J = 7.9 Hz, 1H, Ar), 7.53 (s, 1H, Ar), 7.59 - 7.76 (m, 2H, Ar), 9.01 (t,
J = 5.8 Hz, 1H, NH), 10.98 (s, 1H, NH);
13C NMR (DMSO-
d6)
δ 22.51, 31.20, 42.44, 47.12, 51.56, 114.81, 117.71, 118.12, 122.07, 122.91, 127.07,
127.33, 130.35, 131.60, 142.38, 143.92, 148.78, 167.67, 167.92, 170.98, 172.85; LCMS:
MH
+ = 427, 429; Anal. Calcd. for C
21H
19ClN
4O
4+ 0.2 H
2O: C, 58.73; H, 4.32; N, 13.05; Found: C, 58.68; H, 4.26; N, 12.80.
5.55 Isoindolin compounds (Example disclosed for reference only)
[0330]

[0331] The isoindolin compounds shown above are made as shown in Scheme 2.

[0332] Step 1: DIPEA (2.0 mmol, 2 eq.) is added to a stirred mixture of 3-(5-aminomethyl-1-oxo-1,3-dihydro-isoindol-2-yl)-piperidine-2,6-dione
methanesulfonate
3 (0.26 g, 1.0 mmol) and an appropriate nitrophenylisothiocyanate
4 (1.1 mmol) in acetonitrile (10 mL) under nitrogen. The mixture is stirred at RT for
12 hrs. 1N HCl solution (10 mL) is added and the solid formed is filtered, washed
with additional water, and dried to provide substituted 1-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-3-(2-nitrophenyl)thiourea
5.
[0333] Step 2: To a stirred mixture this intermediate
5 (1.0 mmol) in EtOH (10 mL) is added a solution of sodium dithionite (1.74 g, 10 mmol)
in water (10 mL). The mixture is heated to 60 °C for 1 hr. The reaction mixture is
concentrated to dryness to yield compound
6, which is used directly in the next step without further purification.
[0334] Step 3: Compound
6 is suspended in DMF and EDCI (0.19 g, 1.0 mmol) is added. The mixture is stirred
overnight and the reaction mixture is quenched by addition of acetic acid (2 mL).
The crude product is purified using preparative HPLC to provide a compound 7.
5.56 Isoindoline Compounds (Examples disclosed for reference only)
[0335] According to the procedures provided in Sections 5.34-5.35, 1-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-3-(2-hydroxy-4,5-dimethylphenyl)urea
are prepared starting from 4,5-dimethyl-2-nitrophenol.
[0336] General Procedure A: A mixture of the appropriate carboxylic acid starting material (2.0 mmol) and CDI
(0.32 g, 2.0 mmol) in DMF (30 mL) is stirred for 2 hours at 40°C, and then 3-(5-(aminomethyl)-1-oxoisoindolin-2-yl)piperidine-2,6-dione
methanesulfonate (0.74 g, 2.0 mmol) is added, and stirring proceeds for 24 hours.
The mixture is evaporated under vacuum and the residue is purified by preparative
HPLC.
[0337] Using the
General Procedure A, the following compounds are prepared:

N-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-2-(3-(morpholinomethyl)phenyl)acetamide;

N-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-2-(3-((4-methylpiperazin-1-yl)methyl)phenyl)acetamide;

N-((2-(2,6-dioxopiperidin-3-yl)-l-oxoisoindolin-5-yl)methyl)-2-(4-methyl-3-(morpholinomethyl)phenyl)acetamide;

2-(4-chloro-3-(morpholinomethyl)phenyl)-N-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)acetamide;

N-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-2-(3-(2-morpholinoethyl)phenyl)acetamide;

N-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-2-(4-methyl-3-(2-morpholinoethyl)phenyl)acetamide;

2-(4-chloro-3-(2-morpholinoethyl)phenyl)-N-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)acetamide;

N-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-2-(4-(morpholinomethyl)phenyl)acetamide;

N-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-2-(4-((4-methylpiperazin-1-yl)methyl)phenyl)acetamide;

N-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-2-(3-methyl-4-(morpholinomethyl)phenyl)acetamide;

2-(3-chloro-4-(morpholinomethyl)phenyl)-N-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)acetamide;

N-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-2-(4-(2-morpholinoethyl)phenyl)acetamide;

N-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-2-(3-methyl-4-(2-morpholinoethyl)phenyl)acetamide;

2-(3-chloro-4-(2-morpholinoethyl)phenyl)-N-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)acetamide;

N-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-2-(4-(2-morpholinoethoxy)phenyl)acetamide;

N-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-2-(3-methyl-4-(2-morpholinoethoxy)phenyl)acetamide;

2-(3-chloro-4-(2-morpholinoethoxy)phenyl)-N-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)acetamide;

N-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-2-(3-(2-morpholinoethoxy)phenyl)acetamide;

N-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-2-(4-methyl-3-(2-morpholinoethoxy)phenyl)acetamide;

2-(4-chloro-3-(2-morpholinoethoxy)phenyl)-N-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)acetamide;

N-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-4-(morpholinomethyl)benzamide;

N-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-4-((4-methylpiperazin-1-yl)methyl)benzamide;

N-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-3-methyl-4-(morpholinomethyl)benzamide;

3-chloro-N-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-4-(morpholinomethyl)benzamide;

N-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-4-(2-morpholinoethyl)benzamide;

N-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-3-methyl-4-(2-morpholinoethyl)benzamide;

3-chloro-N-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-4-(2-morpholinoethyl)benzamide;

N-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-3-(morpholinomethyl)benzamide;

N-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-3-((4-methylpiperazin-1-yl)methyl)benzamide;

N-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-4-methyl-3-(morpholinomethyl)benzamide;

4-chloro-N-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-3-(morpholinomethyl)benzamide;

N-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-3-(2-morpholinoethyl)benzamide;

N-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-4-methyl-3-(2-morpholinoethyl)benzamide;

4-chloro-N-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindofin-5-yl)methyl)-3-(2-morpholinoethyl)benzamide;

N-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-4-(2-morpholinoethoxy)benzamide;

N-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-3-methyl-4-(2-morpholinoethoxy)benzamide;

3-chloro-N-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-4-(2-morpholinoethoxy)benzamide;

N-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-3-(2-morpholinoethoxy)benzamide;

N-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-4-methyl-3-(2-morpholinoethoxy)benzamide;
and

4-chloro-N-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-3-(2-morpholinoethoxy)benzamide.
5.57 3-[5-(6-Chloro-4-oxo-4H-quinazolin-3-ylmethyl)-1-oxo-1,3-dihydro-isoindol-2-yl]-piperidine-2,6-dione (Example disclosed for reference only)
[0338]

[0339] 2-Amino-5-chloro-
N-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1
H-isoindol-5-ylmethyl]-benzamide (250 mg, 0.586 mmol) was suspended in trimethyl orthoformate
(3 mL) and the mixture was heated to 85 °C overnight. Formic acid (100 L) was added
to the reaction and the mixture was heated for 1 hr. Water was added to the reaction
resulting in a white precipitate. The solid was filtered, washed with additional water,
and dried in a vacuum oven to provide the product as a white solid (140 mg, 55% yield):
HPLC: Waters Symmetry C
18, 5 µm, 3.9 x 150 mm, 1 mL/min, 240 nm, 30/70 CH
3CN/0.1% H
3PO
4, 7.20 min (97.9 %); mp: 308-310 °C;
1H NMR (DMSO-
d6)
δ 1.90 - 2.08 (m, 1H, CHH), 2.37 (qd,
J = 4.3, 13.2 Hz, 1H, CHH), 2.59 (d,
J = 17.6 Hz, 1H, CHH), 2.78 - 3.03 (m, 1H, CHH), 4.23 - 4.35 (m, 1H, NCHH), 4.38 -
4.51 (m, 1H, NCHH), 5.10 (dd,
J= 5.0, 13.1 Hz, 1H, NCH), 5.33 (s, 2H, NCH
2), 7.52 (d,
J = 7.7 Hz, 1H, Ar), 7.59 (s, 1H, Ar), 7.67 - 7.80 (m, 2H, Ar), 7.88 (dd,
J = 2.5, 8.7 Hz, 1H, Ar), 8.09 (d,
J = 2.3 Hz, 1H, Ar), 8.66 (s, 1H, NCHN), 10.98 (s, 1H, CONH);
13C NMR (DMSO-
d6)
δ 22.45, 31.17, 47.13, 49.20, 51.59, 122.68, 122.91, 123.23, 125.11, 127.56, 129.59,
131.14, 131.50, 134.59, 140.45, 142.59, 146.68, 148.47, 159.20, 167.63, 170.92, 172.82;
LCMS: MH
+ = 437, 439; Anal. Calcd. for C
22H
17ClN
4O
4+ 0.4 H
2O: C, 59.51; H, 4.04; N, 12.62; 7.98, Cl; Found: C, 59.29; H, 3.66; N, 12.39; 7.82,
Cl.
5.58 6-Chloro-3-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-1H-quinazoline-2,4-dione (Example disclosed for reference only)
[0340]

[0341] To a slurry of 2-amino-5-chloro-
N-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1
H-isoindol-5-ylmethyl]-benzamide (250 mg, 0.586 mmol) in acetonitrile (5 mL) was added
DIPEA (0.204 mL, 1.171 mmol), followed by phosgene (0.62 mL, 1.171 mmol). The reaction
mixture was warmed up to 60 °C. After 4 hrs, additional phosgene was added (0.31 mL)
and stirring was continued for 1 hr, at which time LC-MS indicated reaction completion.
The reaction slurry was quenched with dilute HCl, and the white solid was collected
by filtration and washed with additional water and copious Et
2O. The cake was dried in a vacuum oven to afford the product as a white solid (185
mg, 84% yield): HPLC: Waters Symmetry C
18, 5 µm, 3.9 x 150 mm, 1 mL/min, 240 nm, 30/70 CH
3CN/0.1% H
3PO
4, 7.97 min (97.6 %); mp: 358-360 °C;
1H NMR (DMSO-
d6)
δ 1.88
- 2.07 (m, 1H, CHH), 2.24 - 2.45 (m, 1H, CHH), 2.53 - 2.68 (m, 1H, CHH), 2.79 - 3.01
(m, 1H, CHH), 4.17 - 4.36 (m, 1H, CHH), 4.36 - 4.54 (m, 1H, CHH), 5.09 (dd,
J = 4.9, 13.2 Hz, 1H, CH), 5.19 (s, 2H, PhCH
2), 7.24 (d,
J = 8.7 Hz, 1H, Ar), 7.47 (d,
J = 7.9 Hz, 1H, Ar), 7.54 (s, 1H, Ar), 7.68 (d,
J = 7.7 Hz, 1H, Ar), 7.74 (dd,
J = 2.4, 8.8 Hz, 1H, Ar), 7.89 (d,
J = 2.3 Hz, 1H, Ar), 10.98 (s, 1H, NH), 11.72 (s, 1H, NH);
13C NMR (DMSO-
d6)
δ 22.46, 31.18, 43.50, 47.12, 51.58, 115.11, 117.49, 122.29, 122.96, 126.37, 126.54,
127.29, 130.65, 135.04, 138.38, 141.17, 142.39, 149.90, 161.08, 167.79, 170.94, 172.83.
LCMS: MH = 453, 455; Anal. Calcd. for C
22H
17ClN
4O
5 + 0.3 H
2O: C, 57.66; H, 3.87; N, 12.23; Cl, 7.74; Found: C, 57.60; H, 3.90; N, 11.97; Cl,
7.72.
5.59 [2-(2,6-Dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-carbamic acid
4-chloro-3-methyl-phenyl ester (Example disclosed for reference only)
[0342]

[0343] To the solution of para-nitro-phenyl chloroformate (1000 mg, 0.5 mmol) in CH
3CN (5 mL), was added the CH
3CN solution (5 mL) of 4-chloro-m-cresol (71.3 mg, 0.5 mmol) and DIPEA (0.24 mL, 1.5
mmol) dropwise at room temperature. The mixture was stirred at room temperature for
10 min. 3-(5-Aminomethyl-1-oxo-1,3-dihydro-isoindol-2-yl)-piperidine-2,6-dione methanesulfonic
acid salt (0.37 g, 1 mmol) was added followed by additional DIPEA (0.24 mL, 1.5 mmol).
The mixture was stirred overnight at ambient temperature. The mixture was then filtered.
The resulted solid was purified on silica gel column eluted using methanol and methylene
chloride to give [2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-carbamic
acid 4-chloro-3-methylphenyl ester as a white solid (30 mg, 14% yield). HPLC: Waters
Symmetry C
18, 5µm, 3.9 x 150 mm, 1 mL/min, 240 nm, 50/50, CH
3CN/0.1% H
3PO
4,: t
R =3.0 min (97%); mp 225-227 °C;
1H NMR (DMSO-d
6) δ 1.91 - 2.08 (m, 1H, CHH), 2.32 (s, 3H, CH
3), 2.40 (dd,
J = 4.3, 12.8 Hz, 1H, CHH), 2.63 (br. s., 1H, CHH), 2.83 - 3.02 (m, 1H, CHH), 4.22
- 4.56 (m, 4H, CH
2, CH
2), 5.12 (dd,
J = 5.1, 13.2 Hz, 1H, NCH), 7.01 (dd,
J = 2.3, 8.7 Hz, 1H, Ar), 7.17 (d,
J = 2.3 Hz, 1H, Ar), 7.40 (d,
J = 8.7 Hz, 1H, Ar), 7.46 (d,
J = 7.4 Hz, 1H, Ar), 7.55 (s, 1H, Ar), 7.72 (d,
J = 7.9 Hz, 1H, Ar), 8.47 (t,
J = 6.0 Hz, 1H, NH), 10.99 (s, 1H, NH).
13C NMR (DMSO-d
6) δ 19.52, 22.49, 31.20, 44.04, 47.15, 51.59, 121.03, 122.04, 123.04, 124.43, 127.01,
129.33, 130.58, 136.54, 142.45, 143.35, 145.35, 149.62, 154.43, 167.87, 170.99, 172.86;
LC-MS: 442; Anal Calcd for C
22H
20ClN
3O
6C, 59.80; H, 4.56; N, 9.51; Found: C, 59.74; H, 4.45; N, 9.58.
5.60 1-[1-(3,4-Dichloro-phenyl)-ethyl]-3-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-urea (Example disclosed for reference only)
[0344]

[0345] A stirred mixture of 3-(5-aminomethyl-1-oxo-1,3-dihydro-isoindol-2-yl)-piperidine-2,6-dione
methane sulfonate (0.37 g, 1.00 mmol) and CDI (0.18 g, 1.10 mmol) in
N,
N-dimethylformamide (10 mL) was heated to 40 °C under nitrogen overnight. 1-(3,4-Dichloro-phenyl)-ethylamine
(0.19 g, 1.00 mmol) was then added and the mixture was heated at 40 °C overnight.
Water (60 mL) was added, solid precipitated, was filtered, washed with water (30 mL)
and was purified by preparative HPLC (gradient: CH
3CN/H
2O: 10/90 for 5 min, to 100/0 in 10 min, 100/0 for 5 min). Solvent was evaporated and
the residue was triturated in ether (20 mL) for 1 h. The product was then isolated
by filtration and dried
in vacuo to give 1-[1-(3,4-Dichloro-phenyl)-ethyl]-3-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-urea
as a white solid (0.084 g, 17% yield): HPLC: Waters Symmetry C
18, 5µm, 3.9 x 150 mm, 1 mL/min, 240 nm, gradient: CH
3CN/0.1% H
3PO
4: 10/90 to 90/10 in 10 min, 90/10 (5 min): 8.57 min (96.68%); mp: 200-202 °C;
1H NMR (DMSO-d
6) δ 1.32 (d,
J = 7.0 Hz, 3H, CH
3), 1.88 - 2.13 (m, 1H, CHH), 2.28 - 2.47 (m, 1H, CHH), 2.55 - 2.70 (m, 1H, CHH), 2.79
- 3.04 (m, 1H, CHH), 4.18 - 4.55 (m, 4H, CH
2, CH
2), 4.74 (quin,
J = 7.0 Hz, 1H, CH), 5.11 (dd,
J = 4.9, 13.2 Hz, 1H, CH), 6.51 (s, 1H, NH), 6.66 (d,
J = 7.9 Hz, 1H, NH), 7.20 - 7.46 (m, 3H, Ar), 7.48 - 7.62 (m, 2H, Ar), 7.65 (d,
J = 7.7 Hz, 1H, Ar), 10.99 (s, 1H, NH);
13C NMR (DMSO-d
6) δ 22.51, 22.80, 31.20, 42.86, 47.07, 48.18, 51.55, 121.61, 122.81, 126.28, 126.70,
127.78, 128.79, 130.14, 130.38, 130.77, 142.26, 145.29, 147.41, 157.20, 167.95, 170.98,
172.85; LCMS: MH = 489, 491; Anal Calcd for C
23H
22N
4O
4Cl
2 + 0.9 H
2O: C, 54.64, H, 4.75, N, 11.08; Found: C, 54.27, H, 4.66, N, 10.98.
5.61 1-(3-Chloro-4-(2-(2-methoxyethoxy)ethoxy)phenyl)-3-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)urea (Example disclosed for reference only)
[0346]

[0347] 3-(5-aminomethyl-1-oxo-1,3-dihydro-isoindol-2-yl)-piperidine-2,6-dione methanesulfonate
(1.11 g, 3.0 mmol) and 1,1'-Carbonyldiimidazole (535 mg, 3.3 mmol) were suspended
in dry DMF (20 mL) and the mixture was stirred at rt for 24 h. While stirring, a portion
of the reaction mixture (6.7 mL), ∼1 mmol) was transferred to a vial containing 3-chloro-4-(2-(2-methoxyethoxy)ethoxy)aniline
hydrochloride (310 mg, 1.1 mmol). The resulting mixture was stirred at rt overnight
and the reaction progress was monitored by LCMS. After 48 h, additional 3-chloro-4-(2-(2-methoxyethoxy)ethoxy)aniline
hydrochloride (56 mg, 0.2 mmol) was transferred to the reaction mixture and stirring
continued for another 24 h. The reaction mixture was acidified with 1 N HCl and water
was added with stirring until a precipitate formed. The solid was collected by filtration,
suction dried, then dissolved in DMF and purified using C-18 preparatory HPLC to give
1-(3-chloro-4-(2-(2-methoxyethoxy)ethoxy)phenyl)-3-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)urea
as a white solid (390 mg, 72% yield): HPLC:Waters Symmetry C
18, 5 µm, 3.9 x 150 mm, 1 ml/min, 240 nm, 30/70 CH
3CN/0.1% H
3PO
4, 5.40 min (99.8%); mp: 188-190 °C;
1H NMR (DMSO-d
6) δ 1.92 - 2.10 (m, 1H, CHH), 2.29 - 2.47 (m, 1H, CHH), 2.60 (dd,
J = 2.2, 15.4 Hz, 1H, CHH), 2.82 - 3.01 (m, 1H, CHH), 3.25 (s, 3H, CH
3), 3.46 (dd,
J = 3.7, 5.6 Hz, 2H, CH
2), 3.61 (dd,
J = 3.8, 5.7 Hz, 2H, CH
2), 3.73 (d,
J = 4.7 Hz, 2H, CH
2), 4.09 (t,
J = 4.9 Hz, 2H, CH
2), 4.24 - 4.55 (m, 4H, CH
2, CH
2), 5.11 (dd,
J = 5.1, 13.2 Hz, 1H, CH), 6.77 (t,
J = 5.9 Hz, 1H, NH), 7.04 (d,
J = 9.1 Hz, 1H, Ar), 7.19 (dd,
J = 2.5, 9.0 Hz, 1H, Ar), 7.38 - 7.49 (m, 1H, Ar), 7.51 (s, 1H, Ar), 7.64 (d,
J = 2.6 Hz, 1H, Ar), 7.69 (d,
J = 7.7 Hz, 1H, Ar), 8.65 (s, 1H, NH), 10.98 (s, 1H, NH);
13C NMR (DMSO-d
6) δ 22.51, 31.20, 42.80, 47.12, 51.56, 58.05, 68.80, 68.85, 69.84, 71.29, 114.66,
117.58, 119.49, 121.24, 121.88, 122.91, 126.89, 130.29, 134.53, 142.38, 144.84, 148.40,
155.25, 167.95, 171.01, 172.86; LCMS: MH = 545, 547; Anal Calcd for C
26H
29ClN
4O
7 + 0.4 H
2O: C, 56.55; H, 5.44; N, 10.15. Found: C, 56.64; H, 5.34; N, 10.15.
5.62 Assays
5.62.1 TNFα Inhibition Assay in PMBC
[0348] Peripheral blood mononuclear cells (PBMC) from normal donors are obtained by Ficoll
Hypaque (Pharmacia, Piscataway, NJ, USA) density centrifugation. Cells are cultured
in RPMI 1640 (Life Technologies, Grand Island, NY, USA) supplemented with 10% AB+
human serum (Gemini Bio-products, Woodland, CA, USA), 2 mM L-glutamine, 100 U/ml penicillin,
and 100 µg/mL streptomycin (Life Technologies).
[0349] PBMC (2 x 10
5 cells) are plated in 96-well flat-bottom Costar tissue culture plates (Coming, NY,
USA) in triplicate. Cells are stimulated with LPS (from Salmonella abortus equi, Sigma
cat.no. L-1887, St.Louis, MO, USA) at 1 ng/mL final in the absence or presence of
compounds. Compounds provided herein are dissolved in DMSO (Sigma) and further dilutions
are done in culture medium immediately before use. The final DMSO concentration in
all assays can be about 0.25%. Compounds are added to cells 1 hr before LPS stimulation.
Cells are then incubated for 18-20 hrs at 37 °C in 5% CO
2, and supernatants are then collected, diluted with culture medium and assayed for
TNFα levels by ELISA (Endogen, Boston, MA, USA). IC
50s are calculated using non-linear regression, sigmoidal dose-response, constraining
the top to 100% and bottom to 0%, allowing variable slope (GraphPad Prism v3.02).
5.62.2 IL-2 and MIP-3α Production by T Cells
[0350] PBMC are depleted of adherent monocytes by placing 1 x 10
8 PBMC in 10 mL complete medium (RPMI 1640 supplemented with 10% heat-inactivated fetal
bovine serum, 2 mM L-glutamine, 100 U/mL penicillin, and 100 µg/mL streptomycin) per
10 cm tissue culture dish, in 37 °C, 5% CO
2 incubator for 30-60 min. The dish is rinsed with medium to remove all non-adherent
PBMC. T cells are purified by negative selection using the following antibody (Pharmingen)
and Dynabead (Dynal) mixture for every 1 x 10
8 non-adherent PBMC: 0.3 mL Sheep anti-mouse IgG beads, 15 µL anti-CD16, 15 µL anti-CD33,
15 µL anti-CD56, 0.23 mL anti-CD19 beads, 0.23 mL anti-HLA class II beads, and 56
µL anti-CD 14 beads. The cells and bead/antibody mixture is rotated end-over-end for
30-60 min at 4 °C. Purified T cells are removed from beads using a Dynal magnet. Typical
yield is about 50% T cells, 87-95% CD3
+ by flow cytometry.
[0351] Tissue culture 96-well flat-bottom plates are coated with anti-CD3 antibody OKT3
at 5 µg/mL in PBS, 100 µL per well, incubated at 37 °C for 3-6 hrs, then washed four
times with complete medium 100 µL/well just before T cells are added. Compounds are
diluted to 20 times of final in a round bottom tissue culture 96-well plate. Final
concentrations are about 10 µM to about 0.00064 µM. A 10 mM stock of compounds provided
herein is diluted 1:50 in complete for the first 20x dilution of 200 µM in 2% DMSO
and serially diluted 1:5 into 2% DMSO. Each compound is added at 10 µL per 200 µL
culture, to give a final DMSO concentration of 0.1%. Cultures are incubated at 37
°C, 5% CO
2 for 2-3 days, and supernatants analyzed for IL-2 and MIP-3α by ELISA (R&D Systems).
IL-2 and MIP-3α levels are normalized to the amount produced in the presence of an
amount of a compound provided herein, and EC
50s are calculated using non-linear regression, sigmoidal dose-response, constraining
the top to 100% and bottom to 0%, allowing variable slope (GraphPad Prism v3.02).
5.62.3 Cell Proliferation Assay
[0352] Cell lines Namalwa, MUTZ-5, and UT-7 are obtained from the Deutsche Sammlung von
Mikroorganismen und Zellkulturen GmbH (Braunschweig, Germany). The cell line KG-1
is obtained from the American Type Culture Collection (Manassas, VA, USA). Cell proliferation
as indicated by
3H-thymidine incorporation is measured in all cell lines as follows.
[0353] Cells are plated in 96-well plates at 6,000 cells per well in media. The cells are
pretreated with compounds at about 100, 10, 1, 0.1, 0.01, 0.001, 0.0001 and 0 µM in
a final concentration of about 0.25% DMSO in triplicate at 37 °C in a humidified incubator
at 5% CO
2 for 72 hrs. One microcurie of
3H-thymidine (Amersham) is then added to each well, and cells are incubated again at
37 °C in a humidified incubator at 5% CO
2 for 6 hrs. The cells are harvested onto UniFilter GF/C filter plates (Perkin Elmer)
using a cell harvester (Tomtec), and the plates are allowed to dry overnight. Microscint
20 (Packard) (25 µL/well) is added, and plates are analyzed in TopCount NXT (Packard).
Each well is counted for one min. Percent inhibition of cell proliferation is calculated
by averaging all triplicates and normalizing to the DMSO control (0 % inhibition).
Each compound is tested in each cell line in three separate experiments. Final IC
50s are calculated using non-linear regression, sigmoidal dose-response, constraining
the top to 100% and bottom to 0%, allowing variable slope. (GraphPad Prism v3.02).
5.62.4 Immunoprecipitation and Immunoblot
[0354] Namalwa cells are treated with DMSO or an amount of a compound provided herein for
1 hr, then stimulated with 10 U/mL of Epo (R&D Systems) for 30 min. Cell lysates are
prepared and either immunoprecipitated with Epo receptor Ab or separated immediately
by SDS-PAGE. Immunoblots are probed with Akt, phospo-Akt (Ser473 or Thr308), phospho-Gab1
(Y627), Gab1, IRS2, actin, and IRF-1 Abs and analyzed on a Storm 860 Imager using
ImageQuant software (Molecular Dynamics).
5.62.5 Cell Cycle Analysis
[0355] Cells are treated with DMSO or an amount of a compound provided herein overnight.
Propidium iodide staining for cell cycle is performed using CycleTEST PLUS (Becton
Dickinson) according to manufacturer's protocol. Following staining, cells are analyzed
by a FACSCalibur flow cytometer using ModFit LT software (Becton Dickinson).
5.62.6 Apoptosis Analysis
[0356] Cells are treated with DMSO or an amount of a compound provided herein at various
time points, then washed with annexin-V wash buffer (BD Biosciences). Cells are incubated
with annexin-V binding protein and propidium iodide (BD Biosciences) for 10 min. Samples
are analyzed using flow cytometry.
5.62.7 Luciferase Assay
[0357] Namalwa cells are transfected with 4 µg of AP1-luciferase (Stratagene) per 1 x 10
6 cells and 3 µL Lipofectamine 2,000 (Invitrogen) reagent according to manufacturer's
instructions. Six hours post-transfection, cells are treated with DMSO or an amount
of a compound provided herein. Luciferase activity is assayed using luciferase lysis
buffer and substrate (Promega) and measured using a luminometer (Turner Designs).
5.62.8 Anti-Proliferation Assays
[0358] Day 1: The cells are seeded to 96-well plate with 50ul/well in 10%FBS RPMI (w/ Glutamine,
w/o pen-strip) for overnight. The following cells are used:
Colorectal cancer cell: Colo 205 3200cells/well; positive control irinotecan
Pancreatic cancer cell: BXPC-3 1200cells/well; positive control gemcitabine
Prostate cancer cell: PC3 1200cells/well; positive control docetaxel
Breast cancer cell: MDA-MB-231 2400cells/well; positive control paclitaxel
[0359] Day 2: The compounds are serially diluted from 0.00001 µm∼10 µm (or 0.000001∼1 µM) with
50 µl/well (of 2x) and added to the plates in duplicate with relative positive control.
The plates were then incubated at 37°C for 72 hours.
[0360] Day 5: The results are detected by CellTiter Glo method. 100 µl/well of CellTiter Glo reagent
is added to the plates and incubated for 10 minutes at room temperature, and then
analyzed on the Top Count reader. The IC
50 of each compound is typically based on the result of two or more individually experiments.
5.63 TNFα Inhibition
[0361] The properties of certain compounds provided herein in ibhibiting TNFα were assessed
using procedures substantially similar to those described in Section 6.62.1 above.
Tested compounds included: 1-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1
H-isoindol-5-ylmethyl]-3-(4-pyridin-4-ylmethyl-phenyl)-urea; 1-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1
H-isoindol-5-ylmethyl]-3-(4-hydroxymethyl-phenyl)-urea; 1-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1
H-isoindol-5-ylmethyl]-3-[3-(2-methyl-2H-pyrazol-3-yl)-phenyl]-urea; 1-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1
H-isoindol-5-ylmethyl]-3-[3-(2-methyl-imidazol-1-yl)-phenyl]-urea; 1-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1
H-isoindol-5-ylmethyl]-3-[4-(4-methyl-4H-[1,2,4]triazol-3-yl)-phenyl]-urea; 1-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1
H-isoindol-5-ylmethyl]-3-[3-(2-methyl-thiazol-4-yl)-phenyl]-urea; 1-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1
H-isoindol-5-ylmethyl]-3-[3-(1-methyl-1H-pyrazol-3-yl)-phenyl]-urea; 1-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-3-(3-(morpholinomethyl)phenyl)urea;
1-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindofin-5-yl)methyl)-3-(4-methyl-3-nitrophenyl)urea;
1-(3-amino-4-methylphenyl)-3-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)urea;
1-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1
H-isoindol-5-ylmethyl]-3-(3-phenoxy-phenyl)-urea; 1-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-3-(4-nitrophenyl)urea;
N-(4-{3-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1
H-isoindol-5-ylmethyl]-ureido}-phenyl)-acetamide; 1-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-3-(3-hydroxy-4-methylphenyl)urea;
1-[3-(1H-benzoimidazol-2-yl)-4-chloro-phenyl]-3-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-urea;
1-(4-
tert-butyl-cyclohexyl)-3-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1
H-isoindol-5-ylmethyl]-urea; 1-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1
H-isoindol-5-ylmethyl]-3-(4-methyl-cyclohexyl)-urea; 1-(6-chloro-pyridin-3-yl)-3-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-urea;
1-[4-(2,4-difluoro-phenyl)-thiazol-2-yl]-3-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-urea;
and 2,2-dimethyl-propionic acid 3-{5-[3-(3-chloro-4-methyl-phenyl)-ureidomethyl]-1-oxo-1,3-dihydro-isoindol-2-yl}-2,6-dioxo-piperidin-1-ylmethyl
ester. The IC
50 values for all of the tested compounds were determined to be in the range of about
0.2 to 300 nM.
5.64 Antiproliferation - Namalwa
[0362] The antiproliferation of certain compounds provided herein were assessed using Namalwa
cells by following procedures substantially similar to those described in Section
5.62.3 above. The tested compounds included: 1-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1
H-isoindol-5-ylmethyl]-3-(4-pyridin-4-ylmethyl-phenyl)-urea; 1-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1
H-isoindol-5-ylmethyl]-3-(4-hydroxymethyl-phenyl)-urea; 1-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1
H-isoindol-5-ylmethyl]-3-[3-(2-methyl-2H-pyrazol-3-yl)-phenyl]-urea; 1-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1
H-isoindol-5-ylmethyl]-3-[3-(2-methyl-imidazol-1-yl)-phenyl]-urea; 1-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1
H-isoindol-5-ylmethyl]-3-[3-(2-methyl-thiazol-4-yl)-phenyl]-urea; 1-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1
H-isoindol-5-ylmethyl]-3-[3-(1-methyl-1H-pyrazol-3-yl)-phenyl]-urea; 1-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-3-(3-(morpholinomethyl)phenyl)urea;
1-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-3-(4-methyl-3-nitrophenyl)urea;
1-(3-amino-4-methylphenyl)-3-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)urea;
1-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1
H-isoindol-5-ylmethyl]-3-(3-phenoxy-phenyl)-urea; 1-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-3-(4-nitrophenyl)urea;
1-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1
H-isoindol-5-ylmethyl]-3-[3-(3-methyl-[1,2,4]oxadiazol-5-yl)-phenyl]-urea; 1-(4-
tert-butylcyclohexyl)-3-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1
H-isoindol-5-ylmethyl]-urea; 1-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1
H-isoindol-5-ylmethyl]-3-(4-methyl-cyclohexyl)-urea; and 2,2-dimethyl-propionic acid
3-{5-[3-(3-chloro-4-methyl-phenyl)-ureidomethyl]-1-oxo-1,3-dihydro-isoindol-2-yl}-2,6-dioxo-piperidin-1-ylmethyl
ester. The IC
50 values for all of the tested compounds were determined to be in the range of about
0.02 to 40 nM.
5.65 Antiproliferation - PC3
[0363] The antiproliferation of certain compounds provided herein were assessed using PC3
cells by following procedures substantially similar to those described in Section
5.62.8 above. The tested compounds included: 1-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1
H-isoindol-5-ylmethyl]-3-(4-pyridin-4-ylmethyl-phenyl)-urea; 1-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1
H-isoindol-5-ylmethyl]-3-[3-(2-methyl-2H-pyrazol-3-yl)-phenyl]-urea; 1-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1
H-isoindol-5-ylmethyl]-3-[3-(2-methyl-thiazol-4-yl)-phenyl]-urea; 1-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1
H-isoindol-5-ylmethyl]-3-[3-(1-methyl-1H-pyrazol-3-yl)-phenyl]-urea; 1-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-3-(3-(morpholinomethyl)phenyl)urea;
1-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-3-(4-methyl-3-nitrophenyl)urea;
1-(3-amino-4-methylphenyl)-3-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)urea;
1-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1
H-isoindol-5-ylmethyl]-3-(3-phenoxy-phenyl)-urea; 1-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-3-(4-nitrophenyl)urea;
1-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-3-(3-hydroxy-4-methylphenyl)urea;
1-[3-(1H-benzoimidazol-2-yl)-4-chloro-phenyl]-3-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-urea;
1-(4-
tert-butyl-cyclohexyl)-3-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1
H-isoindol-5-ylmethyl]-urea; 1-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1
H-isoindol-5-ylmethyl]-3-(4-methyl-cyclohexyl)-urea; 1-(6-chloro-pyridin-3-yl)-3-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-urea;
1-[4-(2,4-difluorophenyl)-thiazol-2-yl]-3-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-urea;
2,2-dimethyl-propionic acid 3-{5-[3-(3-chloro-4-methyl-phenyl)-ureidomethyl]-1-oxo-1,3-dihydro-isoindol-2-yl}-2,6-dioxo-piperidin-1-ylmethyl
ester; and 1-[1-(3,4-dichloro-phenyl)-ethyl]-3-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-urea.
The IC
50 values for all of the tested compounds were determined to be in the range of about
0.001 to 0.74 µM.
5.66 Antiproliferation - BxPC3
[0364] The antiproliferation of certain compounds provided herein were assessed using BxPC3
cells by following procedures substantially similar to those described in Section
5.62.8 above. The tested compounds included: 1-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1
H-isoindol-5-ylmethyl]-3-(4-pyridin-4-ylmethyl-phenyl)-urea; 1-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1
H-isoindol-5-ylmethyl]-3-[3-(2-methyl-2H-pyrazol-3-yl)-phenyl]-urea; 1-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1
H-isoindol-5-ylmethyl]-3-[3-(2-methyl-thiazol-4-yl)-phenyl]-urea; 1-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1
H-isoindol-5-ylmethyl]-3-[3-(1-methyl-1H-pyrazol-3-yl)-phenyl]-urea; 1-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-3-(3-(morpholinomethyl)phenyl)urea;
1-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-3-(4-methyl-3-nitrophenyl)urea;
1-(3-amino-4-methylphenyl)-3-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)urea;
1-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1
H-isoindol-5-ylmethyl]-3-(3-phenoxy-phenyl)-urea; 1-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-3-(4-nitrophenyl)urea;
1-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-3-(3-hydroxy-4-methylphenyl)urea;
1-[3-(1H-benzoimidazol-2-yl)-4-chloro-phenyl]-3-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-urea;
1-(4-
tert-butyl-cyclohexyl)-3-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1
H-isoindol-5-ylmethyl]-urea; 1-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1
H-isoindol-5-ylmethyl]-3-(4-methyl-cyclohexyl)-urea; 1-[4-(2,4-difluoro-phenyl)-thiazol-2-yl]-3-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-urea;
2,2-dimethyl-propionic acid 3-{5-[3-(3-chloro-4-methyl-phenyl)-ureidomethyl]-1-oxo-1,3-dihydro-isoindol-2-yl{-2,6-dioxo-piperidin-1-ylmethyl
ester; and 1-[1-(3,4-dichloro-phenyl)-ethyl]-3-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-urea.
The IC
50 values for all of the tested compounds were determined to be in the range of about
0.01 to 0.94 µM.
5.67 Antiproliferation - MDAMB321
[0365] The antiproliferation of certain compounds provided herein were assessed using MDAMB321
cells by following procedures substantially similar to those described in Section
5.62.8 above. The tested compounds included: 1-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1
H-isoindol-5-ylmethyl]-3-(4-pyridin-4-ylmethyl-phenyl)-urea; 1-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1
H-isoindol-5-ylmethyl]-3-[3-(2-methyl-2H-pyrazol-3-yl)-phenyl]-urea; 1-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1
H-isoindol-5-ylmethyl]-3-[3-(2-methyl-thiazol-4-yl)-phenyl]-urea; 1-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1
H-isoindol-5-ylmethyl]-3-[3-(1-methyl-1H-pyrazol-3-yl)-phenyl]-urea; 1-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-3-(3-(morpholinomethyl)phenyl)urea;
1-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-3-(4-methyl-3-nitrophenyl)urea;
1-(3-amino-4-methylphenyl)-3-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)urea;
1-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1
H-isoindol-5-ylmethyl]-3-(3-phenoxy-phenyl)-urea; 1-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-3-(4-nitrophenyl)urea;
1-[2-(2,6-Dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1
H-isoindol-5-ylmethyl]-3-[3-(3-methyl-[1,2,4]oxadiazol-5-yl)-phenyl]-urea; 1-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-3-(3-hydroxy-4-methylphenyl)urea;
1-[3-(1H-benzoimidazol-2-yl)-4-chloro-phenyl]-3-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-urea;
1-(4-
tert-butylcyclohexyl)-3-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1
H-isoindol-5-ylmethyl]-urea; 1-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1
H-isoindol-5-ylmethyl]-3-(4-methyl-cyclohexyl)-urea; 1-(6-chloro-pyridin-3-yl)-3-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-urea;
1-[4-(2,4-difluoro-phenyl)-thiazol-2-yl]-3-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-urea;
2,2-dimethyl-propionic acid 3-{5-[3-(3-chloro-4-methyl-phenyl)-ureidomethyl]-1-oxo-1,3-dihydro-isoindol-2-yl}-2,6-dioxo-piperidin-1-ylmethyl
ester; and 1-[1-(3,4-dichloro-phenyl)-ethyl]-3-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-urea.
The IC
50 values for all of the tested compounds were determined to be in the range of about
0.4 to 380 nM.
5.68 Antiproliferation - Colo205
[0366] The antiproliferation of certain compounds provided herein were assessed using Colo205
cells by following procedures substantially similar to those described in Section
5.62.8 above. The tested compounds included: 1-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1
H-isoindol-5-ylmethyl]-3-(4-pyridin-4-ylmethyl-phenyl)-urea; 1-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1
H-isoindol-5-ylmethyl]-3-[3-(2-methyl-2H-pyraaol-3-yl)-phenyl]-urea; 1-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1
H-isoindol-5-ylmethyl]-3-[3-(2-methyl-thiazol-4-yl)-phenyl]-urea; 1-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1
H-isoindol-5-ylmethyl]-3-[3-(1-methyl-1H-pyrazol-3-yl)-phenyl]-urea; 1-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-3-(3-(morpholinomethyl)phenyl)urea;
1-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-3-(4-methyl-3-nitrophenyl)urea;
1-(3-amino-4-methylphenyl)-3-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)urea;
1-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1
H-isoindol-5-ylmethyl]-3-(3-phenoxy-phenyl)-urea; 1-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-3-(4-nitrophenyl)urea;
1-[2-(2,6-Dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1
H-isoindol-5-ylmethyl]-3-[3-(3-methyl-[1,2,4]oxadiazol-5-yl)-phenyl]-urea; 1-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-3-(3-hydroxy-4-methylphenyl)urea;
1-[3-(1H-benzoimidazol-2-yl)-4-chloro-phenyl]-3-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-urea;
1-(4-
tert-butylcyclohexyl)-3-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1
H-isoindol-5-ylmethyl]-urea; 1-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1
H-isoindol-5-ylmethyl]-3-(4-methyl-cyclohexyl)-urea; 1-(6-chloro-pyridin-3-yl)-3-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-urea;
1-[4-(2,4-difluoro-phenyl)-thiazol-2-yl]-3-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-urea;
2,2-dimethyl-propionic acid 3-{5-[3-(3-chloro-4-methyl-phenyl)-ureidomethyl]-1-oxo-1,3-dihydro-isoindol-2-yl}-2,6-dioxo-pipendin-1-ylmethyl
ester; and 1-[1-(3,4-dichloro-phenyl)-ethyl]-3-[2-(2,6-dioxo-piperidin-3-yl)-1-oxo-2,3-dihydro-1H-isoindol-5-ylmethyl]-urea.
The IC
50 values for all of the tested compounds were determined to be in the range of about
0.15 to 130 nM.
[0367] The examples set forth above are provided to give those of ordinary skill in the
art with a complete disclosure and description of how to make and use the claimed
embodiments.